|
1
|
Lipoprotein Drug Delivery Vehicles for Cancer: Rationale and Reason. Int J Mol Sci 2019; 20:ijms20246327. [PMID: 31847457 PMCID: PMC6940806 DOI: 10.3390/ijms20246327] [Citation(s) in RCA: 21] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/06/2019] [Revised: 11/26/2019] [Accepted: 12/04/2019] [Indexed: 12/11/2022] Open
Abstract
Lipoproteins are a family of naturally occurring macromolecular complexes consisting amphiphilic apoproteins, phospholipids, and neutral lipids. The physiological role of mammalian plasma lipoproteins is to transport their apolar cargo (primarily cholesterol and triglyceride) to their respective destinations through a highly organized ligand-receptor recognition system. Current day synthetic nanoparticle delivery systems attempt to accomplish this task; however, many only manage to achieve limited results. In recent years, many research labs have employed the use of lipoprotein or lipoprotein-like carriers to transport imaging agents or drugs to tumors. The purpose of this review is to highlight the pharmacologic, clinical, and molecular evidence for utilizing lipoprotein-based formulations and discuss their scientific rationale. To accomplish this task, evidence of dynamic drug interactions with circulating plasma lipoproteins are presented. This is followed by epidemiologic and molecular data describing the association between cholesterol and cancer.
Collapse
|
|
2
|
Li W, Fu J, Ding Y, Liu D, Jia N, Chen D, Hu H. Low density lipoprotein-inspired nanostructured lipid nanoparticles containing pro-doxorubicin to enhance tumor-targeted therapeutic efficiency. Acta Biomater 2019; 96:456-467. [PMID: 31260821 DOI: 10.1016/j.actbio.2019.06.051] [Citation(s) in RCA: 18] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/04/2019] [Revised: 06/20/2019] [Accepted: 06/26/2019] [Indexed: 02/01/2023]
Abstract
Inefficient tumor accumulation and controlling drug release at the tumor site are two major obstacles limiting the antitumor efficacy of nanoparticle delivery systems. Inspired by the biological structure and function of low-density lipoprotein (LDL), a pH-sensitive ApoB-100/Oleic acid-DOX/NLC (AODN) nanoparticle based on nanostructured lipid carrier (NLC) was prepared in this study. The biological composition of ApoB-containing NLC nanoparticles is similar to that of LDL, which can effectively increase the cycle time and targeting efficiency of nanoparticles. Meantime, the doxorubicin prodrug strategy was used to increase the drug loading of the nanoparticles and achieve drug-sensitive release. In vitro results indicated that AODN nanoparticles can cause more drugs to be phagocytosed by LDL receptor-mediated endocytosis, thus showing high cytotoxicity in 4T1 cells. In vivo experiments have shown that pH-sensitive AODN nanoparticles can cause more drugs to accumulate in the tumor site, reducing systemic toxicity and effectively inhibiting orthotopic breast cancer. These data provide strong evidence that the strategy of combining bionics and prodrug technology provides a new approach to improving the efficiency of chemotherapy drugs in cancer treatment. STATEMENT OF SIGNIFICANCE: Inefficient tumor accumulation and controlling drug release at the tumor site are two major obstacles limiting the antitumor efficacy of nanoparticle delivery systems. Inspired by low density lipoprotein, a pH-sensitive ApoB-100/oleic acid-DOX/NLC (AODN) nanoparticle based on nanostructured lipid carrier (NLC) was prepared. Its biological composition is similar to that of LDL, which can effectively increase the cycle time and targeting efficiency of drugs. Then, the doxorubicin prodrug strategy was used to increase the drug loading of the nanoparticles and achieve drug-sensitive release. AODN nanoparticles can effectively inhibit tumor by effectively accumulating at tumor site and controlling release. The strategy of combining bionics and prodrug technology provides a new approach to improving the efficiency of chemotherapy drugs in cancer treatment.
Collapse
Affiliation(s)
- Wenpan Li
- Department of Pharmaceutics, School of Pharmacy, Shenyang Pharmaceutical University, No. 103, Wenhua Road, Shenyang 110016, PR China
| | - Jia Fu
- Department of Pharmaceutics, School of Pharmacy, Shenyang Pharmaceutical University, No. 103, Wenhua Road, Shenyang 110016, PR China
| | - Ying Ding
- Department of Pharmaceutics, School of Pharmacy, Shenyang Pharmaceutical University, No. 103, Wenhua Road, Shenyang 110016, PR China
| | - Dan Liu
- Department of Pharmaceutics, School of Pharmacy, Shenyang Pharmaceutical University, No. 103, Wenhua Road, Shenyang 110016, PR China
| | - Nan Jia
- Department of Pharmaceutics, School of Pharmacy, Shenyang Pharmaceutical University, No. 103, Wenhua Road, Shenyang 110016, PR China
| | - Dawei Chen
- Department of Pharmaceutics, School of Pharmacy, Shenyang Pharmaceutical University, No. 103, Wenhua Road, Shenyang 110016, PR China.
| | - Haiyang Hu
- Department of Pharmaceutics, School of Pharmacy, Shenyang Pharmaceutical University, No. 103, Wenhua Road, Shenyang 110016, PR China.
| |
Collapse
|
|
3
|
Sobot D, Mura S, Rouquette M, Vukosavljevic B, Cayre F, Buchy E, Pieters G, Garcia-Argote S, Windbergs M, Desmaële D, Couvreur P. Circulating Lipoproteins: A Trojan Horse Guiding Squalenoylated Drugs to LDL-Accumulating Cancer Cells. Mol Ther 2017; 25:1596-1605. [PMID: 28606375 PMCID: PMC5498828 DOI: 10.1016/j.ymthe.2017.05.016] [Citation(s) in RCA: 32] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/27/2016] [Revised: 05/23/2017] [Accepted: 05/23/2017] [Indexed: 11/30/2022] Open
Abstract
Selective delivery of anticancer drugs to rapidly growing cancer cells can be achieved by taking advantage of their high receptor-mediated uptake of low-density lipoproteins (LDLs). Indeed, we have recently discovered that nanoparticles made of the squalene derivative of the anticancer agent gemcitabine (SQGem) strongly interacted with the LDLs in the human blood. In the present study, we showed both in vitro and in vivo that such interaction led to the preferential accumulation of SQGem in cancer cells (MDA-MB-231) with high LDL receptor expression. As a result, an improved pharmacological activity has been observed in MDA-MB-231 tumor-bearing mice, an experimental model with a low sensitivity to gemcitabine. Accordingly, we proved that the use of squalene moieties not only induced the gemcitabine insertion into lipoproteins, but that it could also be exploited to indirectly target cancer cells in vivo.
Collapse
MESH Headings
- Adenocarcinoma/genetics
- Adenocarcinoma/pathology
- Adenocarcinoma/therapy
- Animals
- Antineoplastic Agents/chemistry
- Antineoplastic Agents/pharmacology
- Breast Neoplasms/genetics
- Breast Neoplasms/pathology
- Breast Neoplasms/therapy
- Cell Line, Tumor
- Deoxycytidine/analogs & derivatives
- Deoxycytidine/chemistry
- Deoxycytidine/pharmacology
- Drug Carriers
- Female
- Gene Expression Regulation, Neoplastic
- Humans
- Lipoproteins, LDL/chemistry
- Lipoproteins, LDL/metabolism
- Mice
- Mice, Inbred BALB C
- Mice, Inbred C57BL
- Mice, Nude
- Mice, SCID
- Nanoparticles/administration & dosage
- Nanoparticles/chemistry
- Receptors, LDL/genetics
- Receptors, LDL/metabolism
- Squalene/chemistry
- Tumor Burden/drug effects
- Xenograft Model Antitumor Assays
- Gemcitabine
Collapse
Affiliation(s)
- Dunja Sobot
- Institut Galien Paris-Sud, UMR 8612, CNRS, University Paris-Sud, Université Paris-Saclay, Faculté de Pharmacie, 5 rue Jean-Baptiste Clément, 92296 Châtenay-Malabry Cedex, France
| | - Simona Mura
- Institut Galien Paris-Sud, UMR 8612, CNRS, University Paris-Sud, Université Paris-Saclay, Faculté de Pharmacie, 5 rue Jean-Baptiste Clément, 92296 Châtenay-Malabry Cedex, France
| | - Marie Rouquette
- Institut Galien Paris-Sud, UMR 8612, CNRS, University Paris-Sud, Université Paris-Saclay, Faculté de Pharmacie, 5 rue Jean-Baptiste Clément, 92296 Châtenay-Malabry Cedex, France
| | - Branko Vukosavljevic
- Department of Drug Delivery, Helmholtz Institute for Pharmaceutical Research Saarland, Helmholtz Center for Infection Research, Campus E8 1, 66123 Saarbruecken, Germany
| | - Fanny Cayre
- Institut Galien Paris-Sud, UMR 8612, CNRS, University Paris-Sud, Université Paris-Saclay, Faculté de Pharmacie, 5 rue Jean-Baptiste Clément, 92296 Châtenay-Malabry Cedex, France
| | - Eric Buchy
- Institut Galien Paris-Sud, UMR 8612, CNRS, University Paris-Sud, Université Paris-Saclay, Faculté de Pharmacie, 5 rue Jean-Baptiste Clément, 92296 Châtenay-Malabry Cedex, France
| | - Grégory Pieters
- SCBM, CEA, Université Paris Saclay, LabEx LERMIT, 91191 Gif-sur-Yvette, France
| | | | - Maike Windbergs
- Department of Drug Delivery, Helmholtz Institute for Pharmaceutical Research Saarland, Helmholtz Center for Infection Research, Campus E8 1, 66123 Saarbruecken, Germany; Institute of Pharmaceutical Technology, Buchmann Institute for Molecular Life Sciences, Goethe University, Max-von-Laue-Strasse 15, 60438 Frankfurt am Main, Germany
| | - Didier Desmaële
- Institut Galien Paris-Sud, UMR 8612, CNRS, University Paris-Sud, Université Paris-Saclay, Faculté de Pharmacie, 5 rue Jean-Baptiste Clément, 92296 Châtenay-Malabry Cedex, France
| | - Patrick Couvreur
- Institut Galien Paris-Sud, UMR 8612, CNRS, University Paris-Sud, Université Paris-Saclay, Faculté de Pharmacie, 5 rue Jean-Baptiste Clément, 92296 Châtenay-Malabry Cedex, France.
| |
Collapse
|
|
4
|
Needham D, Arslanagic A, Glud K, Hervella P, Karimi L, Høeilund-Carlsen PF, Kinoshita K, Mollenhauer J, Parra E, Utoft A, Walke P. Bottom up design of nanoparticles for anti-cancer diapeutics: “put the drug in the cancer’s food”. J Drug Target 2016; 24:836-856. [DOI: 10.1080/1061186x.2016.1238092] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/13/2023]
Affiliation(s)
- David Needham
- Department of Mechanical Engineering and Material Science, Duke University, Durham, NC, USA
- Center for Single Particle Science and Engineering (SPSE), University of Southern Denmark, Odense, Denmark
| | - Amina Arslanagic
- Center for Single Particle Science and Engineering (SPSE), University of Southern Denmark, Odense, Denmark
| | - Kasper Glud
- Center for Single Particle Science and Engineering (SPSE), University of Southern Denmark, Odense, Denmark
| | - Pablo Hervella
- Center for Single Particle Science and Engineering (SPSE), University of Southern Denmark, Odense, Denmark
| | - Leena Karimi
- Center for Single Particle Science and Engineering (SPSE), University of Southern Denmark, Odense, Denmark
| | | | - Koji Kinoshita
- Center for Single Particle Science and Engineering (SPSE), University of Southern Denmark, Odense, Denmark
| | - Jan Mollenhauer
- NanoCAN, Institute for Molecular Medicine (IMM), SUND, University of Southern Denmark, Odense, Denmark
| | - Elisa Parra
- Center for Single Particle Science and Engineering (SPSE), University of Southern Denmark, Odense, Denmark
| | - Anders Utoft
- Center for Single Particle Science and Engineering (SPSE), University of Southern Denmark, Odense, Denmark
| | - Prasad Walke
- Center for Single Particle Science and Engineering (SPSE), University of Southern Denmark, Odense, Denmark
| |
Collapse
|
|
5
|
Almer G, Mangge H, Zimmer A, Prassl R. Lipoprotein-Related and Apolipoprotein-Mediated Delivery Systems for Drug Targeting and Imaging. Curr Med Chem 2016; 22:3631-51. [PMID: 26180001 PMCID: PMC5403973 DOI: 10.2174/0929867322666150716114625] [Citation(s) in RCA: 30] [Impact Index Per Article: 3.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/27/2014] [Revised: 06/19/2015] [Accepted: 07/13/2015] [Indexed: 01/27/2023]
Abstract
The integration of lipoprotein-related or apolipoprotein-targeted nanoparticles as pharmaceutical carriers opens new therapeutic and diagnostic avenues in nanomedicine. The concept is to exploit the intrinsic characteristics of lipoprotein particles as being the natural transporter of apolar lipids and fat in human circulation. Discrete lipoprotein assemblies and lipoprotein-based biomimetics offer a versatile nanoparticle platform that can be manipulated and tuned for specific medical applications. This article reviews the possibilities for constructing drug loaded, reconstituted or artificial lipoprotein particles. The advantages and limitations of lipoproteinbased delivery systems are critically evaluated and potential future challenges, especially concerning targeting specificity, concepts for lipoprotein rerouting and design of innovative lipoprotein mimetic particles using apolipoprotein sequences as targeting moieties are discussed. Finally, the review highlights potential medical applications for lipoprotein-based nanoparticle systems in the fields of cardiovascular research, cancer therapy, gene delivery and brain targeting focusing on representative examples from literature.
Collapse
Affiliation(s)
| | | | | | - Ruth Prassl
- Institute of Biophysics, Medical University of Graz, Harrachgasse 21/6, A-8010 Graz, Austria.
| |
Collapse
|
|
6
|
Sapsford KE, Algar WR, Berti L, Gemmill KB, Casey BJ, Oh E, Stewart MH, Medintz IL. Functionalizing nanoparticles with biological molecules: developing chemistries that facilitate nanotechnology. Chem Rev 2013; 113:1904-2074. [PMID: 23432378 DOI: 10.1021/cr300143v] [Citation(s) in RCA: 851] [Impact Index Per Article: 70.9] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/06/2023]
Affiliation(s)
- Kim E Sapsford
- Division of Biology, Department of Chemistry and Materials Science, Office of Science and Engineering Laboratories, U.S. Food and Drug Administration, Silver Spring, Maryland 20993, United States
| | | | | | | | | | | | | | | |
Collapse
|
|
7
|
Abstract
Targeted delivery of anticancer drugs is one of the most actively pursued goals in anticancer chemotherapy. Serum proteins such as transferrin, albumin, and low-density lipoprotein (LDL) offer promise for the selective delivery of antineoplastic agents due to their accumulation in tumor tissue. Uptake of these proteins in solid tumors is mediated by a number of factors, including an increased metabolic activity of tumors, an enhanced vascular permeability of tumor blood vessels for circulating macromolecules, and a lack of a functional lymphatic drainage system in tumor tissue. At the tumor site, transferrin, low-density lipoprotein, and albumin are taken up by the tumor cell through receptor-mediated and fluid phase endocytosis, respectively. Serum protein conjugates can be designed to release the bound antitumor drug after cellular uptake of the drug conjugate. This review covers the diagnostic evidence for tumor accumulation of serum proteins and the design, development, and biological evaluation of drug conjugates with transferrin, albumin, and low-density lipoprotein.
Collapse
Affiliation(s)
- F Kratz
- Department of Medical Oncology, Clinical Research, Tumor Biology Center, Breisacher Strasse 117, Freiburg, Federal Republic of Germany.
| | | |
Collapse
|
|
8
|
Jonkman-De Vries JD, Flora KP, Bult A, Beijnen JH. Pharmaceutical Development of (Investigational) Anticancer Agents for Parenteral Use-A Review. Drug Dev Ind Pharm 2008. [DOI: 10.3109/03639049609108353] [Citation(s) in RCA: 28] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/13/2022]
|
|
9
|
Corbin IR, Li H, Chen J, Lund-Katz S, Zhou R, Glickson JD, Zheng G. Low-density lipoprotein nanoparticles as magnetic resonance imaging contrast agents. Neoplasia 2006; 8:488-98. [PMID: 16820095 PMCID: PMC1601463 DOI: 10.1593/neo.05835] [Citation(s) in RCA: 92] [Impact Index Per Article: 4.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/09/2023] Open
Abstract
Low-density lipoproteins (LDLs) are a naturally occurring endogenous nanoplatform in mammalian systems. These nanoparticles (22 nm) specifically transport cholesterol to cells expressing the LDL receptor (LDLR). Several tumors overexpress LDLRs presumably to provide cholesterol to sustain a high rate of membrane synthesis. Amphiphilic gadolinium (Gd)-diethylenetriaminepentaacetic acid chelates have been incorporated into the LDL to produce a novel LDLR-targeted magnetic resonance imaging (MRI) contrast agent. The number of Gd chelates per LDL particle ranged between 150 and 496 Gd(III). In vitro studies demonstrated that Gd-labeled LDL retained a similar diameter and surface charge as the native LDL particle. In addition, Gd-labeled LDL retained selective cellular binding and uptake through LDLR-mediated endocytosis. Finally, Gd-labeled LDLs exhibited significant contrast enhancement 24 hours after administration in nude mice with human hepatoblastoma G2 xenografts. Thus, Gd-labeled LDL demonstrates potential use as a targeted MRI contrast agent for in vivo tumor detection.
Collapse
Affiliation(s)
- Ian R Corbin
- Department of Radiology, University of Pennsylvania, Philadelphia, PA 19104, USA
| | - Hui Li
- Department of Radiology, University of Pennsylvania, Philadelphia, PA 19104, USA
| | - Juan Chen
- Department of Radiology, University of Pennsylvania, Philadelphia, PA 19104, USA
| | - Sissel Lund-Katz
- Children's Hospital of Philadelphia, Philadelphia, PA 19104, USA
| | - Rong Zhou
- Department of Radiology, University of Pennsylvania, Philadelphia, PA 19104, USA
| | - Jerry D Glickson
- Department of Radiology, University of Pennsylvania, Philadelphia, PA 19104, USA
| | - Gang Zheng
- Department of Radiology, University of Pennsylvania, Philadelphia, PA 19104, USA
| |
Collapse
|
|
10
|
Jiang J, Nilsson-Ehle P, Xu N. Influence of liver cancer on lipid and lipoprotein metabolism. Lipids Health Dis 2006; 5:4. [PMID: 16515689 PMCID: PMC1420303 DOI: 10.1186/1476-511x-5-4] [Citation(s) in RCA: 131] [Impact Index Per Article: 6.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/02/2005] [Accepted: 03/03/2006] [Indexed: 12/13/2022] Open
Abstract
Liver plays a key role in the metabolism of plasma apolipoproteins, endogenous lipids and lipoproteins. Hepatocellular carcinoma (HCC) is one of the most common fatal malignant tumors in China and in other Southeast Asian countries. This has been attributed to the high incidence of hepatitis B infection. Hepatitis B proteins, such as the hepatitis B X protein (HBx) that is large hepatitis B surface protein could regulate transcription of many candidate genes for liver carcinogenesis. It has known that patients who suffered from acute hepatitis B could have lipid disorders such as decreased plasma level of high-density lipoproteins (HDL). Furthermore, aberrations of lipid metabolism are often seen in the chronic hepatitis B infection. Plasma lipid profiles could be changed under HCC. In majority of the reports in HCC, plasma levels of triglycerides (TG), cholesterol, free fatty acids (FFA), HDL, low-density lipoproteins (LDL), lipoprotein (a) (Lp(a)), apolipoprotein AI (apoAI) and apoB were slight to significantly decreased, however, in some cases plasma levels of TG and Lp(a) might be increased. It has been suggested that analysis of plasma levels of lipids, lipoproteins and apolipoproteins in the patients suffered from HCC reflects on the hepatic cellular impairment status. Studies revealed that alterations seen in the plasma levels of lipids, lipoproteins and apolipoproteins reflecting patients' pathologic conditions. Decreased serum levels of cholesterol and apoAI may indicate a poor prognosis. Human leukaemic cells and certain tumor tissues have a higher receptor-mediated uptake of HDL and LDL than the corresponding normal cells or tissues. LDL and HDL have therefore been proposed as a carrier for the water-insoluble anti-cancer agents.
Collapse
Affiliation(s)
- Jingting Jiang
- Section of Clinical Chemistry & Pharmacology, Institute of Laboratory Medicine. Lund University, S-221 85 Lund, Sweden
- Department of Tumor Biological Treatment, The Third Affiliated Hospital, Su Zhou University, Changzhou 213003, China
| | - Peter Nilsson-Ehle
- Section of Clinical Chemistry & Pharmacology, Institute of Laboratory Medicine. Lund University, S-221 85 Lund, Sweden
| | - Ning Xu
- Section of Clinical Chemistry & Pharmacology, Institute of Laboratory Medicine. Lund University, S-221 85 Lund, Sweden
| |
Collapse
|
|
11
|
Lou B, Liao XL, Wu MP, Cheng PF, Yin CY, Fei Z. High-density lipoprotein as a potential carrier for delivery of a lipophilic antitumoral drug into hepatoma cells. World J Gastroenterol 2005; 11:954-9. [PMID: 15742395 PMCID: PMC4250784 DOI: 10.3748/wjg.v11.i7.954] [Citation(s) in RCA: 45] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/06/2023] Open
Abstract
AIM: To investigate the possibility of recombinant high-density lipoprotein (rHDL) being a carrier for delivering antitumoral drug to hepatoma cells.
METHODS: Recombinant complex of HDL and aclacinomycin (rHDL-ACM) was prepared by cosonication of apoproteins from HDL (Apo HDL) and ACM as well as phosphatidylcholine. Characteristics of the rHDL-ACM were elucidated by electrophoretic mobility, including the size of particles, morphology and entrapment efficiency. Binding activity of rHDL-ACM to human hepatoma cells was determined by competition assay in the presence of excess native HDL. The cytotoxicity of rHDL-ACM was assessed by MTT method.
RESULTS: The density range of rHDL-ACM was 1.063-1.210 g/mL, and the same as that of native HDL. The purity of all rHDL-ACM preparations was more than 92%. Encapsulated efficiencies of rHDL-ACM were more than 90%. rHDL-ACM particles were typical sphere model of lipoproteins and heterogeneous in particle size. The average diameter was 31.26±5.62 nm by measure of 110 rHDL-ACM particles in the range of diameter of lipoproteins. rHDL-ACM could bind on SMMC-7721 cells, and such binding could be competed against in the presence of excess native HDL. rHDL-ACM had same binding capacity as native HDL. The cellular uptake of rHDL-ACM by SMMC-7721 hepatoma cells was significantly higher than that of free ACM at the concentration range of 0.5-10 µg/mL (P<0.01). Cytotoxicity of rHDL-ACM to SMMC-7721 cells was significantly higher than that of free ACM at concentration range of less than 5 µg/mL (P<0.01) and IC50 of rHDL-ACM was lower than IC50 of free ACM (1.68 nmol/L vs 3 nmol/L). Compared to L02 hepatocytes, a normal liver cell line, the cellular uptake of rHDL-ACM by SMMC-7721 cells was significantly higher (P<0.01) and in a dose-dependent manner at the concentration range of 0.5-10 μg/mL. Cytotoxicity of the rHDL-ACM to SMMC-7721 cells was significantly higher than that to L02 cells at concentration range of 1-7.5 μg/mL (P<0.01). IC50 for SMMC-7721 cells (1.68 nmol/L) was lower than that for L02 cells (5.68 nmol/L), showing a preferential cytotoxicity of rHDL-ACM for SMMC-7721 cells.
CONCLUSION: rHDL-ACM complex keeps the basic physical and biological binding properties of native HDL and shows a preferential cytotoxicity for SMMC-7721 hepatoma to normal L02 hepatocytes. HDL is a potential carrier for delivering lipophilic antitumoral drug to hepatoma cells.
Collapse
Affiliation(s)
- Bin Lou
- Department of Biochemistry, School of Pharmacy, Fudan University, Shanghai 200032, China
| | | | | | | | | | | |
Collapse
|
|
12
|
Kader A, Pater A. Loading anticancer drugs into HDL as well as LDL has little affect on properties of complexes and enhances cytotoxicity to human carcinoma cells. J Control Release 2002; 80:29-44. [PMID: 11943385 DOI: 10.1016/s0168-3659(01)00536-3] [Citation(s) in RCA: 63] [Impact Index Per Article: 2.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/13/2022]
Abstract
Low density lipoprotein (LDL) has been found to represent a suitable carrier for cytotoxic drugs that may target them to cancer. This study investigated whether very low density lipoprotein (VLDL), LDL and high density lipoprotein (HDL) can be used to effectively incorporate four cytotoxic drugs, 5-fluorouracil (5-FU), 5-iododeoxyuridine (IUdR), doxorubicin (Dox) and vindesine; characterized the complexes; and examined the effect of incorporation on drug cytotoxicity against HeLa cervical and MCF-7 breast carcinoma cells. Significant drug loading was achieved into all three classes of lipoproteins, consistent with the sizes and hydrophobicity of the drugs. The relative loading efficiency was found to be vindesine>IUdR>Dox>5-FU for all three classes of lipoproteins. As shown by electron microscopy (EM), drug incorporation did not affect the size or morphology of the lipoproteins. Differential scanning calorimetry (DSC) showed that drug loading did not significantly change the thermal transition temperature of core lipids in the lipoproteins. The transition enthalpy was changed only for LDL-Dox and LDL-vindesine. The drugs remained stable in the lipoproteins as determined by high performance liquid chromatography (HPLC). EM, DSC and HPLC data suggest that drugs were incorporated into lipoproteins without disrupting their integrity and drugs remained in their stable forms inside lipoproteins. Compared with free drugs in cytotoxicity assays, the IC(50) values of LDL- and HDL-drug complexes were significantly lower (2.4- to 8.6-fold for LDL complexes and 2.5- to 23-fold for HDL complexes). All free or lipoprotein-bound drug formulations were comparably more cytotoxic against MCF-7 than HeLa cells. Upregulating the lipoprotein receptors enhanced, and downregulating them inhibited, the cytotoxicity, indicating the mechanistic involvement of lipoprotein receptor pathways. Complexes of all four drugs with VLDL, in contrast to LDL and HDL, had the same cytotoxicity as the four corresponding free drugs. Our results suggest that further studies are required of the potential of HDL to be a cancer targeting drug carrier.
Collapse
Affiliation(s)
- Abdul Kader
- Faculty of Medicine, Memorial University of Newfoundland, St. John's, NF, Canada A1B 3V6
| | | |
Collapse
|
|
13
|
A synthetic low density lipoprotein particle capable of supporting U937 proliferation in vitro. J Lipid Res 2002. [DOI: 10.1016/s0022-2275(20)30188-7] [Citation(s) in RCA: 24] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/21/2022] Open
|
|
14
|
Owens MD, Baillie G, Halbert GW. Physicochemical properties of microemulsion analogues of low density lipoprotein containing amphiphatic apoprotein B receptor sequences. Int J Pharm 2001; 228:109-17. [PMID: 11576773 DOI: 10.1016/s0378-5173(01)00818-3] [Citation(s) in RCA: 14] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/18/2022]
Abstract
Low density lipoprotein (LDL) has been proposed as a drug targeting vector in cancer chemotherapy, however, research has been limited due to the necessity to isolate material from plasma. In this study, the physicochemical properties of synthetic lipid microemulsions containing an amphiphatic version of the apoprotein B receptor binding sequence have been examined. The effect of peptide sequence length, lipid anchor type and location along with microemulsion lipid composition were investigated via changes in particle size and zeta potential. Size increases were related to the amphiphatic peptides lipophilic portion and too a lesser extent by amino acid sequence length. Two lipophilic anchors, retinoic acid and cholesterol, produced large size increases whilst a single anchor (retinoic acid) did not affect size. The amphiphatic peptide reversed measured zeta potential from negative to positive values in a concentration dependent manner. This was related to peptide structure and could be effected by changes in pH, indicating that the peptide was surface located and responsive to the external environment. Alteration of microemulsion lipid composition also affected physicochemical properties but to a lesser degree than changes in the amphiphatic peptide. These novel systems may represent a useful synthetic alternative to native LDL for a variety of applications.
Collapse
Affiliation(s)
- M D Owens
- Department of Pharmaceutical Sciences, Strathclyde Institute for Biomedical Sciences, University of Strathclyde, 27 Taylor Street, G4 0NR, Glasgow, UK
| | | | | |
Collapse
|
|
15
|
Dorlhiac-Llacer PE, Marquezini MV, Toffoletto O, Carneiro RC, Maranhão RC, Chamone DA. In vitro cytotoxicity of the LDE: daunorubicin complex in acute myelogenous leukemia blast cells. Braz J Med Biol Res 2001; 34:1257-63. [PMID: 11593299 DOI: 10.1590/s0100-879x2001001000004] [Citation(s) in RCA: 11] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/21/2022] Open
Abstract
Acute myelogenous leukemia (AML) blast cells show high-affinity degradation of low-density lipoprotein (LDL), suggesting an increased expression of cellular LDL receptors. LDE is a lipid microemulsion easily synthesized in vitro which is known to mimic the metabolic pathway of LDL. We used LDE as a carrier for daunorubicin and assayed the cytotoxicity of the complex using AML blast cells since RT-PCR analysis showed that AML cells express LDL receptor mRNA. The LDE:daunorubicin complex killed 46.7% of blast cells and 20.2% of normal bone marrow cells (P<0.001; Student t-test). Moreover, this complex destroyed AML blast cells as efficiently as free daunorubicin. Thus, LDE might be a suitable carrier of chemotherapeutic agents targeting these drugs to neoplastic cells and protecting normal tissues.
Collapse
MESH Headings
- Adolescent
- Adult
- Antibiotics, Antineoplastic/pharmacokinetics
- Antibiotics, Antineoplastic/pharmacology
- Child
- Daunorubicin/pharmacokinetics
- Daunorubicin/pharmacology
- Drug Combinations
- Emulsions
- Female
- Humans
- K562 Cells/drug effects
- Leukemia, Myeloid, Acute/drug therapy
- Leukemia, Myeloid, Acute/metabolism
- Leukemia, Myeloid, Acute/pathology
- Lipoproteins, LDL/pharmacokinetics
- Lipoproteins, LDL/pharmacology
- Male
- Receptors, LDL/metabolism
- Reverse Transcriptase Polymerase Chain Reaction
- Tumor Stem Cell Assay
Collapse
Affiliation(s)
- P E Dorlhiac-Llacer
- Departamento de Hematologia e Hemoterapia, Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, Brasil.
| | | | | | | | | | | |
Collapse
|
|
16
|
Masquelier M, Vitols S, Pålsson M, Mårs U, Larsson BS, Peterson CO. Low density lipoprotein as a carrier of cytostatics in cancer chemotherapy: study of stability of drug-carrier complexes in blood. J Drug Target 2000; 8:155-64. [PMID: 10938525 DOI: 10.3109/10611860008996861] [Citation(s) in RCA: 22] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/17/2022]
Abstract
Several solid tumour and leukemia cell types have a higher low density lipoprotein (LDL) uptake than the corresponding normal cells. We are investigating the possibilities to use LDL as a drug carrier to increase the selectivity of antineoplastic drugs in cancer chemotherapy. We have developed a method to incorporate lipophilic cytotoxic agents without interfering with the in vitro and in vivo properties of LDL. In this study, we examined the stability of some drug-LDL complexes in blood and plasma as this is an important prerequisite to achieve a selective therapy. The in vitro dialysis of N-trifluoroacetyl-adriamycin-14-valerat-LDL (AD-32-LDL) against plasma revealed a slow dissociation of the complex. The same method showed a fast and total leakage of paclitaxel from paclitaxel-LDL into the plasma chamber. The dissociation of paclitaxel was confirmed by an autoradiographic study of the distribution of paclitaxel-LDL in tumour-bearing mice. In patients with leukemia the rapid plasma dissociation of AD-32 from LDL illustrated a much higher in vivo instability of this complex. With this method, cholesteryl-linoleate only could be incorporated into LDL in a stable manner as shown by dialysis and autoradiography results. The incorporation of cytotoxic drug derivatives, containing lipophilic anchors, is now under study in order to obtain LDL complexes with better plasma stability.
Collapse
MESH Headings
- Aged
- Aged, 80 and over
- Animals
- Antineoplastic Agents/blood
- Antineoplastic Agents/therapeutic use
- Antineoplastic Agents, Phytogenic/blood
- Antineoplastic Agents, Phytogenic/therapeutic use
- Doxorubicin/analogs & derivatives
- Doxorubicin/blood
- Doxorubicin/therapeutic use
- Drug Carriers
- Female
- Humans
- Leukemia/blood
- Leukemia/drug therapy
- Leukemia, Lymphocytic, Chronic, B-Cell/blood
- Leukemia, Myelogenous, Chronic, BCR-ABL Positive/blood
- Leukemia, Myeloid, Acute/blood
- Lipoproteins, LDL/blood
- Lipoproteins, LDL/therapeutic use
- Male
- Mice
- Mice, Inbred BALB C
- Middle Aged
- Paclitaxel/blood
- Paclitaxel/therapeutic use
Collapse
Affiliation(s)
- M Masquelier
- Department of Medicine, Division of Clinical Pharmacology, Karolinska Institute and Hospital, S-171-76 Stockholm, Sweden.
| | | | | | | | | | | |
Collapse
|
|
17
|
Kader A, Davis PJ, Kara M, Liu H. Drug targeting using low density lipoprotein (LDL): physicochemical factors affecting drug loading into LDL particles. J Control Release 1998; 55:231-43. [PMID: 9795069 DOI: 10.1016/s0168-3659(98)00052-2] [Citation(s) in RCA: 37] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/17/2022]
Abstract
Low density lipoprotein (LDL) has been found suitable as a targeting carrier for cytotoxic drugs. However, higher drug loading into LDL particles without disrupting their native integrity remains a major obstacle. The purpose of this study is to investigate the different physicochemical factors that may affect drug loading and to characterize LDL-drug conjugates. Doxorubicin (Dox) and 3', 5'-O-dipalmitoyl-5-iodo-2'-deoxyuridine (dpIUdR) were used as reference cytotoxic drugs. Drugs were loaded into LDL particles using the dry film method with or without surfactants, liposomal and the direct addition method. The effects of incubation temperature, time and stoichiometry of LDL-drug conjugates on drug loading were investigated. The LDL-drug conjugates were evaluated for their stability and characterized by differential scanning calorimetry (DSC), denatured gel (SDS-PAGE), and electron microscopy (EM). We have suitably incorporated 45+/-10 Dox and 150+/-25 dpIUdR molecules/LDL particle. A seven-fold increase in Dox incorporation was achieved with the liposomal preparation compared to the dry film method. A 4- to 6-h incubation at 37 degreesC was suitable to restore the native structure of LDL particles. No apo B fragmentation of LDL particles was noted on denatured gel. DSC studies showed no change in the Tm of the LDL and the LDL-drug conjugates. An increase in particle size of LDL-dpIUdR, not LDL-Dox was observed in EM compared to the native LDL which may be related to higher incorporation of dpIUdR. The results indicate that physicochemical factors significantly affect drug loading efficiency and may need to be considered to optimize drug incorporation into LDL particles.
Collapse
Affiliation(s)
- A Kader
- School of Pharmacy, Memorial University of Newfoundland, St. John's, NF A1B 3V6, Canada
| | | | | | | |
Collapse
|
|
18
|
Juompan L, Puel J, Fournié GJ, Benoist H. Study of LDL and acetylated LDL endocytosis by mononuclear cells in HIV infection. BIOCHIMICA ET BIOPHYSICA ACTA 1995; 1272:21-8. [PMID: 7545009 DOI: 10.1016/0925-4439(95)00053-7] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/25/2023]
Abstract
Activated lymphocytes have a high level of low density lipoprotein (LDL) uptake as compared to resting lymphocytes, whereas scavenger receptors for acetylated LDL (Ac-LDL) are expressed on limited number of immune cells, i.e., monocytes/macrophages. The endocytosis of LDL and Ac-LDL by mononuclear cells was studied during in vitro and in vivo HIV infection, in order to use LDL and Ac-LDL as carriers of antiviral and/or immunomodulatory drugs towards lymphocytes and monocytes. The uptake of LDL and Ac-LDL was analyzed by cytofluorimetry. LDL endocytosis in PHA/IL2-activated lymphocytes was higher than in resting lymphocytes. In vitro HIV infection of PHA/IL2-activated lymphocytes did not alter the high LDL endocytosis in lymphocytes. CD4+ and CD8+ cells. In a group of 12 symptomatic patients there was no alteration of LDL endocytosis in lymphocytes, CD4 and CD8 lymphocytes. In another group of 23 individuals, the Ac-LDL endocytosis mediated by CD14+ monocytes was unaltered in asymptomatic patients (n = 6) and in some symptomatic patients (n = 6, CD14+ cells > 100/mm3). On the contrary, in other symptomatic patients (n = 11, CD14+ cells < 100/mm3), the number of Ac-LDL+ CD14+ cells decreased, whereas their efficiency of Ac-LDL endocytosis increased as compared to those of other HIV+ patients. In conclusion, the use of lipoproteins as carriers to increase the drug delivery to CD4+ lymphocytes and to CD14+ monocytes can be envisaged, since: (i) the LDL endocytosis was not impaired in CD4 lymphocytes of HIV+ patients, and (ii) the Ac-LDL uptake by monocytes was altered only in some patients of stage IV.
Collapse
MESH Headings
- Antigens, CD/analysis
- Antigens, Differentiation, Myelomonocytic/analysis
- Binding, Competitive
- CD3 Complex/analysis
- CD4-Positive T-Lymphocytes/immunology
- CD4-Positive T-Lymphocytes/metabolism
- CD4-Positive T-Lymphocytes/virology
- CD8-Positive T-Lymphocytes/immunology
- CD8-Positive T-Lymphocytes/metabolism
- CD8-Positive T-Lymphocytes/virology
- Cell Adhesion Molecules
- Cells, Cultured
- Drug Carriers/metabolism
- Endocytosis/physiology
- HIV Infections/immunology
- HIV Infections/metabolism
- HIV-1/physiology
- Humans
- Leukocytes, Mononuclear/immunology
- Leukocytes, Mononuclear/metabolism
- Leukocytes, Mononuclear/virology
- Lipopolysaccharide Receptors
- Lipoproteins, LDL/metabolism
- Lymphocyte Activation
- Monocytes/immunology
- Monocytes/metabolism
- Monocytes/virology
- Receptors, LDL/metabolism
- Receptors, Scavenger
Collapse
Affiliation(s)
- L Juompan
- INSERM U 395, Université Paul Sabatier, Toulouse, France
| | | | | | | |
Collapse
|
|
19
|
De Miguel I, Ioualalen K, Bonnefous M, Peyrot M, Nguyen F, Cervilla M, Soulet N, Dirson R, Rieumajou V, Imbertie L. Synthesis and characterization of supramolecular biovector (SMBV) specifically designed for the entrapment of ionic molecules. BIOCHIMICA ET BIOPHYSICA ACTA 1995; 1237:49-58. [PMID: 7619842 DOI: 10.1016/0005-2736(95)00079-i] [Citation(s) in RCA: 24] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/26/2023]
Abstract
Supramolecular biovectors (SMBV) are nanoparticular drug carriers composed of an internal crosslinked solid core externally grafted with fatty acids and surrounded with a phospholipid layer. We show in this paper that the internal core can be derivatized with anionic ligands such as phosphate in order to allow the efficient entrapment of cationic molecules through a process akin to ion exchange. Synthesis of SMBV involved first a cross linking and derivatization step of polysaccharides followed by a homogenization, a drying and a regioselective acylation step. Acylated polysaccharide cores are thus obtained which can be loaded with drugs and wrapped with a phospholipid layer. The SMBVs obtained are characterized through their size, 20 nm, and their ability to filter through 0.22 microns pore size membrane. Gel permeation chromatography experiments performed with various phospholipid/acylated cores ratios indicate that SMBVs form entities distinct from liposomes and that the optimum phospholipid/acylated cores ratio for this specific type of SMBVs is close to 100%. The supramolecular structure of SMBVs and in particular the spatial proximity between acylated cores and phospholipids is demonstrated through resonance energy transfer experiments. The drug loading capability of SMBVs is illustrated by the preparation of gentamicin and doxorubicin loaded SMBV. The therapeutic potential of SMBVs is then discussed notably in the light of a possible biomimetism with low density lipoproteins (LDL).
Collapse
|
|
20
|
Tokui T, Kuroiwa C, Muramatsu S, Tokui Y, Sasagawa K, Ikeda T, Komai T. Plasma lipoproteins as targeting carriers to tumour tissues after administration of a lipophilic agent to mice. Biopharm Drug Dispos 1995; 16:91-103. [PMID: 7780050 DOI: 10.1002/bdd.2510160204] [Citation(s) in RCA: 12] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/27/2023]
Abstract
We synthesized 14C-warfarin hexadecyl ether (14C-WHE) by addition of a palmityl moiety to the hydroxyl group at the 4-position of 14C-warfarin, a compound known to bind to serum albumin. 14C-WHE preferentially bound to the lipoproteins, low-density lipoprotein (LDL) and high-density lipoprotein (HDL), in mouse plasma both in vitro and in vivo. 14C-Warfarin mainly concentrated in the liver immediately after intravenous administration to mice bearing M5076 sarcoma, and was found at only low concentrations in other tissues including the tumour. 14C-WHE highly distributed to the tumour, adrenal, and spleen, as well as the liver. These tissues coincided with those in which human 125I-LDL was vigorously incorporated. The results indicate that chemical modification of an agent, giving it high lipophilicity, will enable it to bind to lipoproteins after intravenous administration. These modifications raise the possibility of lipoproteins as endogenous targeting carriers into tumour cells, which have high LDL-receptor activity.
Collapse
Affiliation(s)
- T Tokui
- Analytical and Metabolic Research Laboratories, Sankyo Co., Ltd., Tokyo, Japan
| | | | | | | | | | | | | |
Collapse
|
|
21
|
Leppälä J, Kallio M, Nikula T, Nikkinen P, Liewendahl K, Jääskeläinen J, Savolainen S, Gylling H, Hiltunen J, Callaway J. Accumulation of 99mTc-low-density lipoprotein in human malignant glioma. Br J Cancer 1995; 71:383-7. [PMID: 7841057 PMCID: PMC2033577 DOI: 10.1038/bjc.1995.78] [Citation(s) in RCA: 25] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/27/2023] Open
Abstract
Low-density lipoprotein (LDL) uptake in gliomas was studied to find out if LDL has potential as a drug carrier of boron, especially for boron neutron capture therapy. Single photon emission tomography (SPET) was performed 2 h and 20 h after intravenous injection of autologous 99mTc-labelled LDL in four patients with untreated and five patients with recurrent glioma. 99mTc-LDL uptake was compared with the uptake of 99mTc-labelled human serum albumin (HSA), an established blood pool marker. The intra- and peritumoral distributions of radioactivity in the SPET images were not identical for radiolabelled LDL and HSA. The mean LDL tumour to brain ratio, determined from transversal SPET slices at 20 h post injection, was 1.5 in untreated and 2.2 in recurrent gliomas; the corresponding ratios for HSA were 1.6 and 3.4. The brain to blood ratio remained constant at 2 h and 20 h in both types of tumours. These data are not consistent with highly selective, homogeneous uptake of LDL in gliomas. However, the different tumoral distribution and rate of uptake of 99mTc-LDL, as compared with 99mTc-HSA, indicate that the uptake of LDL is different from that of HSA and that further studies on the mechanism of LDL uptake in glioma are warranted.
Collapse
Affiliation(s)
- J Leppälä
- Department of Neurosurgery, University of Helsinki, Finland
| | | | | | | | | | | | | | | | | | | |
Collapse
|
|
22
|
Tokui T, Takatori T, Shinozaki N, Ishigami M, Shiraishi A, Ikeda T, Tsuruo T. Delivery and cytotoxicity of RS-1541 in St-4 human gastric cancer cells in vitro by the low-density-lipoprotein pathway. Cancer Chemother Pharmacol 1995; 36:1-6. [PMID: 7720169 DOI: 10.1007/bf00685724] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/26/2023]
Abstract
RS-1541 is a 13-O-palmitoyl derivative of rhizoxin, an inhibitor of tubulin polymerization. RS-1541 has been shown to bind preferentially to plasma lipoproteins and to exhibit selective and sustained uptake by tumors in mice. To elucidate a mechanism of RS-1541 cytotoxicity, the cellular uptake and the cytotoxicity of a complex of RS-1541 with human low-density lipoprotein (RS-1541/LDL complex) were investigated in cultured St-4 human gastric cancer cells. Both the cellular uptake and the cytotoxicity of the RS-1541/LDL complex were greater in cells with higher LDL-receptor activities than in control cells. Excess amounts of LDL or 1 microM of monensin, a proton ionophore, significantly inhibited both the uptake and the cytotoxicity of the complex. Chloroquine, an inhibitor of lysosomal enzymes, decreased the intracellular level of rhizoxin liberated from RS-1541 and suppressed the cytotoxicity of the RS-1541/LDL complex. However, a detergent-aided solution of RS-1541 showed very low cellular uptake and cytotoxicity, irrespective of the LDL-receptor activities of these cells. These results demonstrate that the RS-1541/LDL complex is incorporated into the cells via the LDL receptor and that it manifests its cytotoxic activity after forming rhizoxin, the original antitumor agent, in the lysosomes.
Collapse
Affiliation(s)
- T Tokui
- Analytical and Metabolic Research Laboratories, Sankyo Co., Ltd., Tokyo, Japan
| | | | | | | | | | | | | |
Collapse
|
|
23
|
Taro T, Yoko T, Michi I, Kazuhiko T, Toshihiko I, Toru K. Targeting of an antitumor agent, RS-1541 (palmitoyl-rhizoxin), via low-density lipoprotein receptor. Int J Pharm 1994. [DOI: 10.1016/0378-5173(94)90250-x] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/27/2022]
|
|
24
|
Bennis F, Favre G, Le Gaillard F, Soula G. Importance of mevalonate-derived products in the control of HMG-CoA reductase activity and growth of human lung adenocarcinoma cell line A549. Int J Cancer 1993; 55:640-5. [PMID: 8406993 DOI: 10.1002/ijc.2910550421] [Citation(s) in RCA: 59] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/30/2023]
Abstract
HMG-CoA reductase catalyzes the synthesis of mevalonate, a crucial intermediate in the biosynthesis of cholesterol and non-sterol isoprenoid compounds essential for cell growth. The HMG-CoA reductase activity of the A549 tumor cell line is higher than that of normal human fibroblasts. This deregulation in mevalonate needs was not due to an alteration in the activated state of the enzyme by short-term regulation. We show that the HMG-CoA reductase in A549 cell line was subject to a multivalent feedback control. A high fraction (40%) of the reductase activity was devoted to non-sterol products. In contrast, normal fibroblasts had only 15-20% of the reductase activity that generated non-sterol products. We also show that cholesterol and at least one of the non-sterol products are necessary for optimal cell growth of A549 cells. Our data strongly suggest that A549 cells produce more non-sterol substances which may be related to increased requirements of mevalonate for upregulated cell growth.
Collapse
Affiliation(s)
- F Bennis
- Laboratoire de Ciblage en Thérapeutique, Biologie de la Cellule Tumorale, UFR des Sciences Pharmaceutiques (Université Paul Sabatier), Toulouse, France
| | | | | | | |
Collapse
|
|
25
|
|
|
26
|
Lundberg B, Hong K, Papahadjopoulos D. Conjugation of apolipoprotein B with liposomes and targeting to cells in culture. BIOCHIMICA ET BIOPHYSICA ACTA 1993; 1149:305-12. [PMID: 8391843 DOI: 10.1016/0005-2736(93)90215-l] [Citation(s) in RCA: 28] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/30/2023]
Abstract
Mixed phospholipid/cholesterol (2:1 molar ratio) liposomes were conjugated with native and acetylated apolipoprotein B (apoB), the protein part of low density lipoprotein (LDL). The objective was to increase the specificity of the cellular uptake of liposomes by utilization of the LDL and scavenger receptor pathways. The method of choice for the conjugation of liposomes with apoB proved to be the detergent solubilization and removal procedure. Two detergents were tested;sodium cholate (NaC) and octyl glucoside (OG). The integrity of the resulting complexes was demonstrated by Sepharose CL-4B gel chromatography and Metrizamide gradient centrifugation. The conjugates showed a good physical stability and the leakiness was only marginally larger than for unconjugated liposomes. The interaction of apoB- and acetyl apoB-liposome conjugates with CV-1 and J774 cells, respectively, was monitored by an encapsulated pH-sensitive fluorophore, pyranine (8-hydroxy-1,3,6-pyrenetrisulfonate (HPTS)). This dye provides means of detecting binding and endocytosis of conjugates in living cells. The internalization was a fast process and about 10-times faster for the OG-conjugates than for the corresponding unconjugated liposomes. The conjugates showed a clear concentration-dependent association of dye with cells, while this was less prominent with liposomes. The uptake was nearly an order of magnitude faster with CV-1 cells than with J774 cells. Acidification of intracellular conjugates proceeded fast during the first 30 min of incubation and reached a minimum value of approx. pH 6 after 3 h. The specificity of binding of apoB-liposome conjugates to CV-1 cells was demonstrated by displacement experiments with native LDL. The results indicate that apoB-liposome conjugates may be used as a delivery vehicle for bioactive subtsances to cells.
Collapse
Affiliation(s)
- B Lundberg
- Department of Biochemistry and Pharmacy, Abo Akademi University, Finland
| | | | | |
Collapse
|
|
27
|
Gueddari N, Favre G, Hachem H, Marek E, Le Gaillard F, Soula G. Evidence for up-regulated low density lipoprotein receptor in human lung adenocarcinoma cell line A549. Biochimie 1993; 75:811-9. [PMID: 8274533 DOI: 10.1016/0300-9084(93)90132-c] [Citation(s) in RCA: 56] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/29/2023]
Abstract
Human lung adenocarcinoma cell line A549 was studied with respect to the metabolism of human low density lipoprotein (LDL) and 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase (HMGR) activity. After incubation in medium containing lipoprotein-deficient serum (LPDS) for 24 h, the A549 cell line expresses a single class of high affinity LDL binding sites (KD at 37 degrees C of 15.1 +/- 0.7 nM and capacity of 118 +/- 2.8 ng/mg cell protein) and an HMGR activity of 111.4 +/- 7 pmol/min/mg cell protein. After binding, the LDLs were internalized and degraded by a common saturable process. The HMGR activity was higher in A549 cells than in fibroblasts but LDL affinity and binding capacity were similar in both cell types. However, in the presence of lipoproteins, A549 cells showed a two-fold higher binding capacity than fibroblasts. When the cells were deprived of cholesterol, the amount of LDLR sites increased but the extent of stimulation was lower in A549 than in fibroblast cells (2.5-fold versus six-fold respectively). This increase was accompanied by a similar increase in the specific LDLR mRNA cellular levels (two-fold versus six-fold respectively). When cells were deprived of exogenous and endogenous cholesterol (biosynthesis blocked by compactin), the binding capacity and the LDLR mRNA levels were yet again increased in A549 cells but not in fibroblasts. Taken together these results suggest that the level of expression of the LDLR is up-regulated in A549 cells compared to fibroblasts.
Collapse
Affiliation(s)
- N Gueddari
- Laboratoire de Ciblage en Thérapeutique, Faculté des Sciences Pharmaceutiques, Toulouse, France
| | | | | | | | | | | |
Collapse
|