Standard neoadjuvant chemotherapy, based on taxanes and anthracyclines, makes conservative treatment of breast cancer possible and it allows for the evaluation of the tumor response in terms of achieving pathological complete response. Whereas hypoxia participates in carcinogenesis, resulting in less differenced tumor cells and poorer prognosis, HIF-1α could be predictive of the tumor response to treatment. Nonetheless, very few studies have evaluated the predictive value of HIF-1α in breast cancer in patients receiving neoadjuvant chemotherapy.

To demonstrate the value of hypoxia-inducible factor-1α (HIF-1α) in predicting response in patients with breast cancer receiving standard neoadjuvant chemotherapy (NAC). Methods: Ninety-five women enrolled in two prospective studies underwent biopsies for the histopathological diagnosis of breast carcinoma before receiving NAC, based on anthracyclines and taxanes. For expression of HIF-1α, EGFR, pAKT and pMAPK, tumor samples were analyzed by immunohistochemistry in tissues microarrays. Standard statistical methods (Pearson chi-square test, Fisher exact test, Kruskal–Wallis test, Mann–Whitney test and Kaplan–Meier method) were used to study the association of HIF-1α with tumor response, survival and other clinicopathologic variables/biomarkers. Results: HIF-1α expression was positive in 35 (39.7%) cases and was significantly associated to complete pathological response (pCR) (p = 0.014). HIF-1α expression was correlated positively with tumor grade (p = 0.015) and Ki-67 expression (p = 0.001) and negativity with progesterone receptors (PR) (p = 0.04) and luminal A phenotype expression (p = 0.005). No correlation was found between HIF-1α expression and EGFR, pAKT and pMAPK. In terms of survival, HIF-1α expression was associated with a significantly shorter disease-free survival (p = 0.013), being identified as an independent prognostic factor in multivariate analysis. Conclusions: Overexpression of HIF-1α is a predictor of pCR and shorter DFS; it would be valuable to confirm these results in prospective studies.

• AKT
• BRET
• Chemoresistance
• ERK1/2
• HGSOC
• Second line non-platinum drugs

• Akt
• Breast cancer
• Drug resistance
• Hypoxic conditions
• Protein kinase B
• Targeted therapy

• HIF-1α
• Shc protein
• T cell
• cell survival
• hypoxia

The hypoxia-inducible transcription factors (HIF)-1/2α are the main oxygen sensors which regulate the adaptation to intratumoral hypoxia. The aim of this study was to assess the role of the HIF proteins in regulating the radiation response of a non-small cell lung cancer (NSCLC) in vitro model. To directly assess the unique and overlapping functions of HIF-1α and HIF-2α, we use CRISPR gene-editing to generate isogenic H1299 non-small cell lung carcinoma cells lacking HIF-1α, HIF-2α or both. We found that in HIF1 knockout cells, HIF-2α was strongly induced by hypoxia compared to wild type but the reverse was not seen in HIF2 knockout cells. Cells lacking HIF-1α were more radiation resistant than HIF2 knockout and wildtype cells upon hypoxia, which was associated with a reduced recruitment of γH2AX foci directly after irradiation and not due to differences in proliferation. Conversely, double-HIF1/2 knockout cells were most radiation sensitive and had increased γH2AX recruitment and cell cycle delay. Compensatory HIF-2α activity in HIF1 knockout cells is the main cause of this radioprotective effect. Under hypoxia, HIF1 knockout cells uniquely had a strong increase in lactate production and decrease in extracellular pH. Using genetically identical HIF-α isoform-deficient cells we identified a strong radiosensitizing of HIF1, but not of HIF2, which was associated with a reduced extracellular pH and reduced glycolysis.

• HIF
• hypoxia
• metabolism
• radiotherapy

• Basic Helix-Loop-Helix Transcription Factors/genetics
• Basic Helix-Loop-Helix Transcription Factors/physiology
• Carcinoma, Non-Small-Cell Lung/genetics
• Carcinoma, Non-Small-Cell Lung/radiotherapy
• Cell Line, Tumor
• Gene Knockout Techniques
• Humans
• Hypoxia-Inducible Factor 1, alpha Subunit/genetics
• Hypoxia-Inducible Factor 1, alpha Subunit/physiology
• Lung Neoplasms/genetics
• Lung Neoplasms/radiotherapy
• Radiation Tolerance/genetics

• Acute Lung Injury/metabolism
• Acute Lung Injury/prevention & control
• Animals
• Cytokines/metabolism
• Escherichia coli
• Flow Cytometry
• Hypoxia-Inducible Factor 1, alpha Subunit/deficiency
• Lipopolysaccharides
• Lung/metabolism
• Lung/pathology
• Male
• Mice
• Mice, Knockout
• Myeloid Cells/metabolism
• Pulmonary Edema
• Spleen/metabolism
• Spleen/pathology

• Chronic myeloid leukemia
• HIF-1α
• p21
• p53

• AKT
• HIF-1
• HNSCC
• NSCLC
• adenocarcinoma
• hypoxia
• squamous cell carcinoma

• Angiogenesis
• Cancer
• Development
• Hypoxia
• Hypoxia-inducible Factor (HIF)
• Vascular Endothelial Growth Factor (VEGF)

• Animals
• Breast Neoplasms/metabolism
• Cell Differentiation/drug effects
• Female
• Human Umbilical Vein Endothelial Cells/cytology
• Human Umbilical Vein Endothelial Cells/drug effects
• Human Umbilical Vein Endothelial Cells/metabolism
• Humans
• Lipopolysaccharide Receptors/metabolism
• Macrophage Colony-Stimulating Factor/pharmacology
• Mice
• Monocytes/cytology
• Monocytes/drug effects
• Monocytes/metabolism
• Receptor, TIE-2/metabolism

• R01 MH067167 NIMH NIH HHS
• R01 HL067167 NHLBI NIH HHS
• R00 CA131552 NCI NIH HHS
• R01 HL109481 NHLBI NIH HHS
• P30 CA016058 NCI NIH HHS
• R25 GM089571 NIGMS NIH HHS

• HIF-1
• cancer metastasis
• cancer therapy

The etiology of inflammatory myofibroblastic tumors (IMTs) is controversial and the prognosis is unpredictable. Previous studies have not investigated the expression of hypoxia-related markers in IMTs.

Between 2002 and 2012, 12 consecutive patients with histologically proven IMTs were enrolled in the study. Immunohistochemistry was used to detect GLUT-1, HIF-1α, PI3K, and p-Akt expression in paraffin-embedded tumor specimens. Associations among GLUT-1, HIF-1α, PI3K, and p-Akt protein expression and clinical parameters were investigated.

The mean duration of follow-up was 52.1 months (range, 11 to 132 months). Six patients had local recurrence. GLUT-1, HIF-1α, PI3K, and p-Akt expression were detected in 41.7%, 50.0%, 33.3%, and 41.7% of patients, respectively. Fisher’s exact test revealed significant correlations between recurrence of IMT and PI3K expression (P = 0.01) and p-Akt expression (P = 0.015). Univariate analyses revealed significant correlations between survival and GLUT-1 expression (P = 0.028), PI3K expression (P = 0.006), and p-Akt expression (P = 0.028). Multivariate analysis did not show a significant relationship between survival and GLUT-1, HIF-1α, PI3K, or p-Akt. Spearman rank correlation analysis showed significant correlations between HIF-1α and PI3K expression (r = 0.707, P = 0.01) and between p-Akt and PI3K expression (r = 0.837, P = 0.001).

Although our results are inconclusive owing to the small sample size, they suggest that PI3K and p-Akt expression may play a role in the recurrence of IMTs of the head and neck.

DDX3 is an RNA helicase that has antiapoptotic properties, and promotes proliferation and transformation. In addition, DDX3 was shown to be a direct downstream target of HIF-1α (the master regulatory of the hypoxia response) in breast cancer cell lines. However, the relation between DDX3 and hypoxia has not been addressed in human tumors. In this paper, we studied the relation between DDX3 and the hypoxic responsive proteins in human breast cancer.

DDX3 expression was investigated by immunohistochemistry in breast cancer in comparison with hypoxia related proteins HIF-1α, GLUT1, CAIX, EGFR, HER2, Akt1, FOXO4, p53, ERα, COMMD1, FER kinase, PIN1, E-cadherin, p21, p27, Transferrin receptor, FOXO3A, c-Met and Notch1. DDX3 was overexpressed in 127 of 366 breast cancer patients, and was correlated with overexpression of HIF-1α and its downstream genes CAIX and GLUT1. Moreover, DDX3 expression correlated with hypoxia-related proteins EGFR, HER2, FOXO4, ERα and c-Met in a HIF-1α dependent fashion, and with COMMD1, FER kinase, Akt1, E-cadherin, TfR and FOXO3A independent of HIF-1α.

In invasive breast cancer, expression of DDX3 was correlated with overexpression of HIF-1α and many other hypoxia related proteins, pointing to a distinct role for DDX3 under hypoxic conditions and supporting the oncogenic role of DDX3 which could have clinical implication for current development of DDX3 inhibitors.

• Adaptor Proteins, Signal Transducing/metabolism
• Adult
• Aged
• Aged, 80 and over
• Breast Neoplasms/enzymology
• Cell Cycle Proteins
• DEAD-box RNA Helicases/genetics
• DEAD-box RNA Helicases/metabolism
• ErbB Receptors/metabolism
• Female
• Forkhead Transcription Factors
• Gene Expression Regulation, Neoplastic/genetics
• Gene Expression Regulation, Neoplastic/physiology
• Humans
• Hypoxia/enzymology
• Hypoxia/metabolism
• Hypoxia-Inducible Factor 1, alpha Subunit/metabolism
• Immunohistochemistry
• In Vitro Techniques
• Male
• Middle Aged
• Proto-Oncogene Proteins c-akt/metabolism
• Receptor, ErbB-2/metabolism
• Receptor, Notch1/metabolism
• Transcription Factors/metabolism

• MC_U130184144 Medical Research Council
• MC_UU_12014/2 Medical Research Council

Protein-tyrosine phosphorylation, which is catalyzed by protein-tyrosine kinase (PTK), plays a pivotal role in a variety of cellular functions related to health and disease. The discovery of the viral oncogene Src (v-Src) and its cellular nontransforming counterpart (c-Src), as the first example of PTK, has opened a window to study the relationship between protein-tyrosine phosphorylation and the biology and medicine of cancer. In this paper, we focus on the roles played by Src and other PTKs in cancer cell-specific behavior, that is, evasion of apoptosis or cell death under stressful extracellular and/or intracellular microenvironments (i.e., hypoxia, anoikis, hypoglycemia, and serum deprivation).

• Animals
• Antineoplastic Agents/metabolism
• Antineoplastic Agents/pharmacology
• Antineoplastic Agents/therapeutic use
• Drug Resistance, Multiple/genetics
• Drug Resistance, Neoplasm/genetics
• Gene Expression Regulation, Neoplastic
• Genes, MDR
• Humans
• Models, Biological
• Multidrug Resistance-Associated Proteins/genetics
• Multidrug Resistance-Associated Proteins/metabolism
• Neoplasm Proteins/genetics
• Neoplasm Proteins/metabolism
• Neoplasms/drug therapy
• Neoplasms/genetics
• Oxidation-Reduction

• R01 CA079085 NCI NIH HHS
• R01 CA089541 NCI NIH HHS
• CA79085 NCI NIH HHS
• CA72404 NCI NIH HHS

Hepatocyte growth factor (HGF), through Met receptor binding, fulfils numerous functions in invasive tumour growth (survival/proliferation, motility, apoptosis), but epigenetic control of gene expression in this process is poorly understood. In HGF-treated breast cancer cells we studied (a) the chemoinvasion towards CXCL12 (ligand of the chemokine-receptor CXCR4) and (b) the mechanistic basis, that is, the transduction pathways that regulate CXCR4-mediated invasion, and the role played by histone deacetylases (HDACs) after blockade with trichostatin A (TSA). In highly invasive and metastatic MDA-MB231 cells HGF had a dual inhibitory effect, reducing spontaneous migration and specific chemoinvasion towards CXCL12, the latter by decreasing CXCR4 transactivation and protein level. After HGF the levels of phosphorylated (therefore active) c-Src and Akt persistently increased, indicating a role of these signal transducers in the HGF-dependent cellular and molecular effects. c-Src wild-type expression vector (Srcwt) increased active c-Src and mimicked the HGF-dependent inhibition of CXCR4 transactivation. Our findings indicate that HDACs participated in the HGF-inhibitory effects. In fact, blockade of HDACs hindered the HGF- and Srcwt-dependent reductions of CXCR4 transactivation and invasiveness, while inhibition of endogenous c-Src was additive with HGF, further reducing specific chemoinvasion. In conclusion, in MDA-MB231 cells HDAC blockade with TSA partly counteracted the HGF-dependent effects through molecular events that included enhancement of the expression of the genes for invasiveness Met and CXCR4 (depending on serum conditions), reduction of endogenous phospho-c-Src/c-Src and phosphoAkt/Akt ratios and triggering of apoptosis. The potential therapeutic use of TSA should take into account the variable aggressiveness of breast carcinoma cells and microenvironment signals such as HGF at the secondary growth site of the tumour. It was interesting that HGF reduced motility and CXCR4 functionality only of MDA-MB231 cells, and not of low-invasive MCF-7 cells, suggesting a mechanism implicated in metastatic cell homing.

• immune-deficient mice
• human stem cells
• mesenchymal stem cells
• hypoxia
• transplantation
• tissue repair

• Animals
• Apoptosis
• Cell Movement
• Humans
• Hypoxia
• Ischemia
• Mesenchymal Stem Cell Transplantation/methods
• Mesenchymal Stem Cells/cytology
• Mice
• Muscles/metabolism
• Oxygen/metabolism
• Proto-Oncogene Proteins c-akt/metabolism
• Proto-Oncogene Proteins c-met/metabolism
• Regeneration
• Signal Transduction

• 2R01-DK53041 NIDDK NIH HHS
• R01 DK053041 NIDDK NIH HHS
• R01-HL073256 NHLBI NIH HHS
• R01 DK061848 NIDDK NIH HHS
• 2R01-DK61848 NIDDK NIH HHS
• R01 HL073256 NHLBI NIH HHS

Expression of the transcription factor hypoxia-inducible factor 1 (HIF-1), which plays a key role in cellular adaptation to hypoxia, was investigated in normal colorectal mucosa (ten), adenomas (61), and carcinomas (23). Tissue samples were analyzed for HIF-1α, its upstream regulators, von Hippel–Lindau factor, AKT, and mammalian target of rapamycin (mTOR) and its downstream targets glucose transporter 1 (GLUT1), carbonic anhydrase IX, stromal-cell-derived factor 1 (SDF-1) by immunohistochemistry. In normal colorectal mucosa, HIF-1α was observed in almost all nuclei of surface epithelial cells, probably secondary to a gradient of oxygenation, as indicated by pimonidazole staining. The same staining pattern was present in 87% of adenomas. In carcinomas, HIF-1α was present predominantly around areas of necrosis (78%). Active AKT and mTOR, were present in all adenomas, carcinomas, and in normal colorectal mucosa. GLUT1 and SDF-1 were present in the normal surface epithelium of all adenoma cases, whereas in the carcinoma GLUT1 was located around necrotic regions and SDF-1 was present in all epithelial cells. In conclusion, HIF-1α appears to be physiologically expressed in the upper part of the colorectal mucosa. The present observations support that upregulation of HIF-1α and its downstream targets GLUT1 and SDF-1 in colorectal adenomas and carcinomas may be due to hypoxia, in close interaction with an active phosphatidylinositol 3-kinases–AKT–mTOR pathway.

• Adult
• Aged
• Aquaporin 1/biosynthesis
• Breast Neoplasms/metabolism
• Cell Movement
• Cell Proliferation
• Female
• Gene Expression Regulation, Neoplastic
• Humans
• Hypoxia
• Hypoxia-Inducible Factor 1/metabolism
• Immunohistochemistry/methods
• Middle Aged
• Neoplasm Metastasis
• Signal Transduction

• Animals
• Basic Helix-Loop-Helix Transcription Factors/physiology
• Blotting, Western
• Caenorhabditis elegans/genetics
• Caenorhabditis elegans/growth & development
• Caenorhabditis elegans/metabolism
• Caenorhabditis elegans Proteins/genetics
• Caenorhabditis elegans Proteins/metabolism
• Cell Hypoxia
• Cells, Cultured
• Deferoxamine/pharmacology
• Gene Expression Regulation
• Genome
• HeLa Cells
• Humans
• Hypoxia-Inducible Factor 1, alpha Subunit/genetics
• Hypoxia-Inducible Factor 1, alpha Subunit/metabolism
• Nuclear Proteins/metabolism
• Procollagen-Proline Dioxygenase/antagonists & inhibitors
• Procollagen-Proline Dioxygenase/metabolism
• RNA, Messenger/metabolism
• RNA, Small Interfering/genetics
• RNA, Small Interfering/metabolism
• Repressor Proteins/genetics
• Repressor Proteins/metabolism
• Response Elements
• Transcription, Genetic
• Transcriptional Activation
• Transfection
• Twist-Related Protein 1/metabolism

• Blotting, Western
• Breast Neoplasms/metabolism
• Breast Neoplasms/pathology
• Electrophoretic Mobility Shift Assay
• Hepatocyte Growth Factor/pharmacology
• Histone Deacetylases/metabolism
• Humans
• Immunoprecipitation
• NF-kappa B/genetics
• NF-kappa B/metabolism
• Neoplasm Invasiveness
• Phosphorylation
• Promoter Regions, Genetic/genetics
• Proto-Oncogene Proteins pp60(c-src)/metabolism
• Receptors, CXCR4/genetics
• Receptors, CXCR4/metabolism
• Transcriptional Activation

AIM: To investigate the implication of the hypoxia inducible factor HIF-1α mRNA in gastric carcinoma and its relation to the expression of vascular endothelial growth factor (VEGF) protein, tumor angiogenesis invasion/metastasis and the patient's survival.

METHODS: In situ hybridization was used to examine expression of HIF-1α mRNA, and immunohistochemical staining was used to examine expression of VEGF protein and CD34 in 118 specimens from patients with gastric carcinoma.

RESULTS: The positive rates of HIF-1α mRNA and VEGF protein were 49.15% and 55.92%, respectively. Positive expressions of HIF-1α and VEGF in stage T₃-T₄ tumors and those with vessel invasion, lymph node metastasis and distant metastasis were dramatically stronger than stage T₁-T₂ cases and those without vessel invasion, lymph node metastasis and distant metastasis. The mean microvascular density (MVD) in stage T₃-T₄ tumors and those with vessel invasion, lymph node metastasis and distant metastasis was significantly higher than stage T₁-T₂ tumors and those without vessel invasion, lymph node metastasis and distant metastasis. The mean MVD in tumors with positive HIF-1α and VEGF expression was significantly higher than that in tumors with negative HIF-1α and VEGF expression. The expression of HIF-1α was positively correlated with VEGF protein. There were positive correlations between MVD and expression of HIF-1α and VEGF. The mean survival time and the 5-year survival rate in cases with positive expression HIF-1α and VEGF and MVD value ≥ 41.5/0.72 mm² were significantly lower than those with negative expression of HIF-1α and VEGF and MVD value < 41.5/0.72 mm².

CONCLUSION: Overexpression of HIF-1α is found in gastric carcinoma. HIF-1α may induce the angiogenesis in gastric carcinoma by upregulating the transcription of VEGF gene, and take part in tumor invasion and metastasis. They can be used as prognostic markers of gastric cancer in clinical practice.

• Gastric carcinoma
• HIF-1α
• Vascular endothelial growth factor
• Microvascular density
• Prognosis

• Adenocarcinoma/blood supply
• Adenocarcinoma/metabolism
• Adenocarcinoma/pathology
• Adult
• Aged
• Carcinoma, Signet Ring Cell/blood supply
• Carcinoma, Signet Ring Cell/metabolism
• Carcinoma, Signet Ring Cell/pathology
• Female
• Gene Expression Regulation, Neoplastic/physiology
• Humans
• Hypoxia-Inducible Factor 1, alpha Subunit/genetics
• Hypoxia-Inducible Factor 1, alpha Subunit/physiology
• Male
• Middle Aged
• Neoplasm Invasiveness/physiopathology
• Neoplasm Metastasis/physiopathology
• Neovascularization, Pathologic/physiopathology
• Prognosis
• RNA, Messenger/metabolism
• Stomach/pathology
• Stomach/physiopathology
• Stomach Neoplasms/blood supply
• Stomach Neoplasms/metabolism
• Stomach Neoplasms/pathology
• Survival Rate
• Up-Regulation
• Vascular Endothelial Growth Factor A/genetics
• Vascular Endothelial Growth Factor A/metabolism

• Acetophenones/pharmacology
• Antioxidants/pharmacology
• Cell Line, Tumor
• DNA/drug effects
• DNA/metabolism
• Dose-Response Relationship, Drug
• Enzyme Inhibitors/pharmacology
• Gene Expression Regulation/drug effects
• Hepatocytes/drug effects
• Hepatocytes/metabolism
• Humans
• Hydrogen Peroxide/metabolism
• Hypoxia-Inducible Factor 1/genetics
• Hypoxia-Inducible Factor 1/metabolism
• Insulin/metabolism
• Insulin/pharmacology
• Myocytes, Cardiac/drug effects
• Myocytes, Cardiac/metabolism
• NADPH Oxidases/antagonists & inhibitors
• NADPH Oxidases/metabolism
• Onium Compounds/pharmacology
• Phosphatidylinositol 3-Kinases/metabolism
• Proline/analogs & derivatives
• Proline/pharmacology
• Promoter Regions, Genetic/drug effects
• Promoter Regions, Genetic/genetics
• Protein Tyrosine Phosphatases/antagonists & inhibitors
• Protein Tyrosine Phosphatases/metabolism
• Reactive Oxygen Species/metabolism
• Reactive Oxygen Species/pharmacology
• Receptor, Insulin/drug effects
• Receptor, Insulin/metabolism
• Signal Transduction/drug effects
• Thiocarbamates/pharmacology
• Time Factors
• Transcription, Genetic/drug effects
• Transfection
• Vanadates/pharmacology