骨桥蛋白shRNA表达载体下调骨桥蛋白表达对卵巢癌细胞侵袭性的影响The effect of shRNA expression vector of osteopontin gene after interfereing protein expression on ovarian cancer cell invasiveness
徐伟;张岚;施薇;
摘要(Abstract)：
目的:构建靶向骨桥蛋白(OPN)的shRNA表达载体,通过RNA干扰(RNAi)下调OPN在卵巢癌细胞HO-8910PM中的表达,观察对卵巢癌细胞侵袭能力的影响,探索卵巢癌基因治疗新途径。方法:将人OPN两条shRNA表达序列克隆到pSilenc-er3.1-H1neo质粒的黏性末端,并对重组质粒进行酶切及序列鉴定。将构建好的重组质粒转染入对数生长期高转移性人卵巢癌细胞株HO-8910PM,Western blot法分析转染后卵巢癌细胞OPN蛋白表达情况,Transwell小室检测细胞的侵袭、迁移情况。结果:OPN-shRNA表达载体克隆构建成功,插入片段测序结果与合成的shRNA结果一致。Western blot结果显示,卵巢癌细胞HO-8910PM转染OPN-shRNA表达载体后,OPN蛋白表达显著降低(P<0.05),卵巢癌细胞的侵袭、迁移能力亦显著降低(P<0.05)。结论:靶向OPN的shRNA表达载体构建成功,可抑制高转移性卵巢癌细胞株OPN表达,并降低细胞侵袭、迁移能力,为研究降低卵巢癌OPN表达对发挥抗肿瘤效应打下基础。
关键词(KeyWords)： 骨桥蛋白;遗传载体;RNA,小分子干扰;RNA干扰;质粒;细胞系,卵巢肿瘤,HO-8910PM
基金项目(Foundation):
作者(Authors): 徐伟;张岚;施薇;
DOI: 10.13283/j.cnki.xdfckjz.2013.02.008
参考文献(References)：
- [1]Senger DR,Wirth DF,Hynes RO.Transformed mammaliancells secrete specific proteins and phosphoproteins[J].Cell,1979,16(4):885-893
- [2]Shevde LA,Das S,Clark DW,et al.Osteopontin:an effec-tor and an effect of tumor metastasis[J].Curr Mol Med,2010,10(1):71-81
- [3]Yang L,Wei L,Zhao W,et al.Down-regulation of os-teopontin expression by RNA interference affects cell pro-liferation and chemotherapy sensitivity of breast cancerMDA-MB-231 cells[J].Mol Med Report,2012,5(2):373-376
- [4]Wai PY,Mi Z,Guo H,et al.Osteopontin silencing bysmall interfering RNA suppresses in vitro and in vivoCT26 murine colon adenocarcinoma metastasis[J].Carci-nogenesis,2005,26(4):741-751
- [5]温敏杰,朱光辉,蔡文松.靶向骨桥蛋白基因的RNA干扰对裸鼠肝癌影响的实验研究[J].临床和实验医学杂志,2008,7(2):1-2
- [6]Coppola D,Szabo M,Boulware D,et al.Correlation of os-teopontin protein expression and pathological stage acrossa wide variety of tumor histologies[J].Clin Cancer Res,2004,10(1 Pt 1):184-190
- [7]张丽丽,邵淑丽,武燕.OPN和B7-H4在上皮性卵巢肿瘤中的表达及意义[J].癌症,2010,29(1):25-29
- [8]Nakae M,Iwamoto I,Fujino T,et al.Preoperative plasmaosteopontin level as a biomarker complementary to carbo-hydrate antigen 125 in predicting ovarian cancer[J].JObstet Gynaecol Res,2006,32(3):309-314
- [9]Song G,Cai QF,Mao YB,et al.Osteopontin promotes o-varian cancer progression and cell survival and increasesHIF-1alpha expression through the PI3-K/Akt pathway[J].Cancer Sci,2008,99(10):1901-1907