孙开来, 110001, 辽宁省沈阳市和平区北二马路92号, 中国医科大学医学遗传学教研室. sunkailai@21cn. com
This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Dong-Mei Hao, Xiu-Ju Sun, Zhi-Hong Zheng, Guang He, Ming-Chao Ma, Kai-Lai Sun, Department of Medical Genetics, Chinese Medical University, Shenyang 110001, Liaoning Province, China
Hui-Mian Xu, Mei-Xian Wang, Department of Oncology, First Affiliated Hospital, Chinese Medical University, Shenyang 110001, Liaoning Province, China
Supported by: the National Major Basic Research Development Program, No. G1998051203.
Correspondence to: Dr. Kai-Lai Sun, Department of Medical Genetics, Chinese Medical University, No. 92, the 2th Northern Road, Heping District, Shenyang 110001, Liaoning Province, China. sunkailai@21cn. com
Received: November 12, 2002 Revised: November 25, 2002 Accepted: November 28, 2002 Published online: January 15, 2003
AIM: To explore molecular mechanism of gastric carcinogenesis, we screened associated genes of gastric dysplasia and further investigated their expression in gastric carcinomas with different stages.
METHODS: Relatively pure dysplasia and normal tissue were procured by manual microdissection, amplified by cDNA-PCR, and then used to carry out forward(dysplasia as tester, normal tissue as driver) and reverse(normal tissue as tester, dysplasia as driver) SSH. Subtracted cDNA fragments were cloned into vector, screened, sequenced, and made homologous analysis. The expression of differentially expressed fragments was detected and verified by Dot hybridization and reverse transcription-PCR.
RESULTS: Two subtracted cDNA libraries were constructed. Twenty-one of 26 sequenced clones were verified to be expressed differentially. It was noted that differentiall expressions of 4 genes(P125, cytochrome c oxidase subunit I, meprin A, acidic calponin)were detected simultaneously in dysplasia, early cancer and advanced cancer.
CONCLUSION: Four new associated genes have been identified in dysplasia. Further studies are necessary to determine their roles in gastric carcinogenesis.
Key Words: N/A
Citation: Hao DM, Sun XJ, Zheng ZH, He G, Ma MC, Xu HM, Wang MX, Sun KL. Screening and expression of associated genes in gastric dysplasia. Shijie Huaren Xiaohua Zazhi 2003; 11(1): 6-9
Zheng ZH, Xun XJ, Qiu GR, Liu YH, Wang MX, Sun KL. E-cadherin gene mutation in precancerous condition、early and advanced stage of gastric cancer.Shijie Huaren Xiaohua Zazhi. 2002;10:153-156.
Cheng SD, Wu YL, Zhang YP, Qiao MM, Guo QS. Abnormal drug accumulation in multidrug resistant gastric carcinoma cells.Shijie Huaren Xiaohua Zazhi. 2001;9:131-134.
Wang B, Shi LC, Zhang WB, Xiao CM, Wu JF, Dong YM. Expression and significance of P16 gene in gastric cancer and its precancerous lesions.Shijie Huaren Xiaohua Zazhi. 2001;9:39-42.
Chen SY, Wang JY, Ji Y, Zhang XD, Zhu CW. Effects of Helicobacter pylori and protein kinase C on gene mutation in gastric cancer and precancerous lesions.Shijie Huaren Xiaohua Zazhi. 2001;9:302-307.
Yao XX, Yin L, Zhang JY, Bai WY, Li YM, Sun ZC. hTERT expression and cellular immunity in gastric cancer and precancerosis.Shijie Huaren Xiaohua Zazhi. 2001;9:508-512.
Guo YQ, Zhu ZH, Li JF. Flow cytometric analysis of apoptosis and proliferation in gastric cancer and precancerous lesion.Shijie Huaren Xiaohua Zazhi. 2000;8:983-987.
Wang DX, Fang DC, Liu WW. Study on alteration of multiple genes in intestinal metaplasia, atypical hyperplasia and gastric cancer.Shijie Huaren Xiaohua Zazhi. 2000;8:855-859.
Zhou YN, Xu CP, Han B, Wang JS, Fang DC, Peng GY, Yang JM, Liu HF. E-cadherin expression correlates with clinicopathological features in gastric carcinoma.Shijie Huaren Xiaohua Zazhi. 2002;10:1121-1124.
Liu HF, Liu WW, Fang DC, Yang SM, Wang RQ. Bax gene expression and its relationship with apoptosis in human gastric carcinoma and precancerous lesions.Shijie Huaren Xiaohua Zazhi. 2000;8:665-668.
He XX, Wang JL, Wu JL, Yuan SY, Ai L. Telomerase expression, H. pylori infection and gastric mucosal carcinogenesis.Shijie Huaren Xiaohua Zazhi. 2000;8:505-508.
He XX, Wang JL, Wu JL, Yuan SY, Ai L. Telomere, cellular DNA content and gastric mucosal carcinogenesis.Shijie Huaren Xiaohua Zazhi. 2000;8:509-512.
Luo ZB, Luo YH, Lu R, Jin HY, Zhang PB, Xu CP. Immunohistochemical study on dendritic cells in gastric mucosa of patients with gastric cancer and precancerous lesions.Shijie Huaren Xiaohua Zazhi. 2000;8:400-402.
Wang RQ, Fang DC, Liu WW. MUC2 gene expression in gastric cancer and preneoplastic lesion tissues.Shijie Huaren Xiaohua Zazhi. 2000;8:285-288.
Mi JQ, Zhang ZH, Shen MC. Significance of CD44v6 protein expression in gastric carcinoma and precancerous lesions.Shijie Huaren Xiaohua Zazhi. 2000;8:156-158.
Wang W, Luo HS, Yu BP. Expression of human telomerase reverse transcriptase gene and c-myc protein in gastric carcinogenesis.Shijie Huaren Xiaohua Zazhi. 2002;10:258-261.
Han P, Liu Y, Cheng J, Wang G, Lu YY, Li K, Li L. The expression of oncogene c-myc upregulated by c-terminally truncated middle surface protein o f hepatitis B virus.Shijie Huaren Xiaohua Zazhi. 2002;10:141-144.
Zeng JX, Wang WL, Luo WJ, Wang ZL. Construction of differentially expressed cDNA library in apoptotic human hepatocellular carcinoma cells with suppression subtractive hybridization.Shijie Huaren Xiaohua Zazhi. 2001;9:1233-1237.
Li C, Liang Y, Wu M, Xu L, Cai W. Telomerase activity analysis of esophageal carcinoma using microdissection-TRAP assay.Chin Med J (Engl). 2002;115:1405-1408.
Li C, Wu M, Liang Y, Xu L, Cai W. [Analysis of telomerase activity in esophageal carcinoma using microdissection telomeric repeat amplification protocol assay].Zhonghua Yi Xue Za Zhi. 2002;82:39-42.
Bowler LD, Hubank M, Spratt BG. Representational difference analysis of cDNA for the detection of differential gene expression in bacteria: development using a model of iron-regulated gene expression in Neisseria meningitidis.Microbiology. 1999;145:3529-3537.
Melià MJ, Bofill N, Hubank M, Meseguer A. Identification of androgen-regulated genes in mouse kidney by representational difference analysis and random arbitrarily primed polymerase chain reaction.Endocrinology. 1998;139:688-695.
Diatchenko L, Lau YF, Campbell AP, Chenchik A, Moqadam F, Huang B, Lukyanov S, Lukyanov K, Gurskaya N, Sverdlov ED. Suppression subtractive hybridization: a method for generating differentially regulated or tissue-specific cDNA probes and libraries.Proc Natl Acad Sci U S A. 1996;93:6025-6030.
Zhang JZ, Huang YW, Liu DM, Yang LT, Han RG, Wang JF, Yue MX. Tissue microarray technology and its preliminary application in large intestinal carcinoma.Shijie Huaren Xiaohua Zazhi. 2002;10:157-160.
Alevizos I, Mahadevappa M, Zhang X, Ohyama H, Kohno Y, Posner M, Gallagher GT, Varvares M, Cohen D, Kim D. Oral cancer in vivo gene expression profiling assisted by laser capture microdissection and microarray analysis.Oncogene. 2001;20:6196-6204.
Kitahara O, Furukawa Y, Tanaka T, Kihara C, Ono K, Yanagawa R, Nita ME, Takagi T, Nakamura Y, Tsunoda T. Alterations of gene expression during colorectal carcinogenesis revealed by cDNA microarrays after laser-capture microdissection of tumor tissues and normal epithelia.Cancer Res. 2001;61:3544-3549.
Yang GP, Ross DT, Kuang WW, Brown PO, Weigel RJ. Combining SSH and cDNA microarrays for rapid identification of differentially expressed genes.Nucleic Acids Res. 1999;27:1517-1523.
Rho J, Altmann CR, Socci ND, Merkov L, Kim N, So H, Lee O, Takami M, Brivanlou AH, Choi Y. Gene expression profiling of osteoclast differentiation by combined suppression subtractive hybridization (SSH) and cDNA microarray analysis.DNA Cell Biol. 2002;21:541-549.
Weber A, Engers R, Nockemann S, Gohr LL, Zur Hausen A, Gabbert HE. Differentially expressed genes in association with in vitro invasiveness of human epithelioid sarcoma.Mol Pathol. 2001;54:324-330.
Tani K, Mizoguchi T, Iwamatsu A, Hatsuzawa K, Tagaya M. p125 is a novel mammalian Sec23p-interacting protein with structural similarity to phospholipid-modifying proteins.J Biol Chem. 1999;274:20505-20512.
Köhler D, Kruse M, Stöcker W, Sterchi EE. Heterologously overexpressed, affinity-purified human meprin alpha is functionally active and cleaves components of the basement membrane in vitro.FEBS Lett. 2000;465:2-7.
Lottaz D, Maurer CA, Hahn D, Büchler MW, Sterchi EE. Nonpolarized secretion of human meprin alpha in colorectal cancer generates an increased proteolytic potential in the stroma.Cancer Res. 1999;59:1127-1133.
Sasaki Y, Yamamura H, Kawakami Y, Yamada T, Hiratsuka M, Kameyama M, Ohigashi H, Ishikawa O, Imaoka S, Ishiguro S. Expression of smooth muscle calponin in tumor vessels of human hepatocellular carcinoma and its possible association with prognosis.Cancer. 2002;94:1777-1786.
Yamamura H, Yoshikawa H, Tatsuta M, Akedo H, Takahashi K. Expression of the smooth muscle calponin gene in human osteosarcoma and its possible association with prognosis.Int J Cancer. 1998;79:245-250.
van Dekken H, Alers JC, Riegman PH, Rosenberg C, Tilanus HW, Vissers K. Molecular cytogenetic evaluation of gastric cardia adenocarcinoma and precursor lesions.Am J Pathol. 2001;158:1961-1967.