应用功能分类基因芯片筛选卵巢癌淋巴结定向高转移细胞系中差异性表达基因Screening and identification for lymphatic metastasis-associated differentially expressed genes in ovarian carcinoma cell lines with directional highly lymphatic metastasis using cDNA microarray
冠潇;李力;黎丹戎;阮和云;尹永硕;张玮;
摘要(Abstract):
目的:在裸鼠体内建立以人卵巢浆液性乳头状腺癌细胞系SKOV3具有淋巴结定向高转移亚克隆第二代SKOV3-pm2、第三代SKOV3-pm3细胞系,应用基因芯片技术比较这3种具有不同淋巴结定向转移能力的细胞亚系中基因表达谱的差异,寻找卵巢癌侵袭转移相关基因。方法:采用Trizol一步法抽提SKOV3、SKOV3-pm2、SKOV3-pm33种细胞总RNA;取等量RNA逆转录合成cDNA用Ampolabeling-LPR(linear polymerase re-action,线性聚合酶反应)法标记的cDNA链做探针,混合后与功能分类基因芯片[OligoTumor Metastasis Microarray Catalog No.OHS-028(人)]杂交,计算机分析后比较3种细胞中差异性表达基因,并应用RT-PCR技术对其中3个表达差异明显的基因进行验证。结果:共筛选出表达差异性基因60个,其中表达上调基因47个(ratio>3),有21个基因在SKOV3-pm2、SKOV3-pm3中共同表达上调;表达下调基因13个(ratio<0.5),有1个基因在SKOV3-pm2、SKOV3-pm3细胞中共同表达下调。结论:基因表达芯片检测与肿瘤淋巴体外转移模型相结合,为肿瘤转移研究提供了新方法、新思路。
关键词(KeyWords): 卵巢肿瘤;功能分类基因芯片;淋巴结定向转移;差异表达基因
基金项目(Foundation): 教育部高等学校博士学科点专项科研基金资助项目(No:20050598002);; 广西自然科学基金资助项目(桂科基No:0575087);; 广西自治区大型仪器协作共用网资助项目
作者(Authors): 冠潇;李力;黎丹戎;阮和云;尹永硕;张玮;
DOI: 10.13283/j.cnki.xdfckjz.2008.01.023
参考文献(References):
- [1]Yoshida BA,Sokoloff MM,Welch DR,et al.Metastasis-suppressor genes:a reviewand perspective on an emergingfield[J].J Natl Cancer Inst,2000,92:1717-1730
- [2]Habermann H,Ray V,Habermann W,et al.Alterations in gap junction protein expression in human benign prostatic hyperplasia and prostate cancer[J].J Urol,2002,167:655-660
- [3]Harada H,Takahashi M.CD44-dependent intracellular and extracellular catabolism of hyaluronic acid by hyaluroni-dase-1and-2[J].J Biol Chem,2007,282:5597-5607
- [4]Besson A,Robbins SM,Yong VW.PTEN/MMAC1/TEP1in signal transduction and tumorigenesis[J].Eur J Bio-chem,1999,263:605-611
- [5]Haier J,Nicolson GL.Time-dependent dephosphorylation through serine/threonine phosphatases is required for sta-ble adhesion of highly and poorly metastatic HT-29colon carcinoma cell lines to collagen[J].Anticancer Res,2000,20:2265-2271
- [6]Kang-Park S,Lee YI,Lee YI.PTENmodulates insulin-like growth factor II(IGF-II)-mediated signaling;the protein phosphatase activity of PTEN downregulates IGF-II ex-pression in hepatoma cells[J].FEBS Lett,2003,545:203-208
- [7]Schondorf T,Ebert MP,Hoffmann J,et al.Hypermethyla-tion of the PTEN gene in ovarian cancer cell lines[J].Cancer Lett,2004,207:215-220
- [8]Kurose K,Zhou XP,Araki T,et al.Frequent loss of PTEN expression is linked to elevated phosphorylated AKT lev-els,but not associated with p27and cyclin D1expression,in primary epithelial ovarian carcinomas[J].Am J Pathol,2001,158:2097-2106
- [9]Helfer B.Boswell BC,Finlay D,et al.Caspase-8promotes cell motility and calpain activity under nonapoptotic condi-tions[J].Cancer Res,2006,66:4273-4278