摘要
目的筛查宫颈癌组织中差异表达蛋白,探讨survivin基因与宫颈癌发生发展的关系。方法利用蛋白阵列方法分析宫颈癌组织与正常宫颈组织中蛋白表达的差异;将体外培养的宫颈癌Hela细胞分为对照组、空质粒组和实验组,转染survivin siRNA表达载体后,利用半定量RT-PCR和Western blotting方法检测各组细胞survivin基因的转录及蛋白表达情况;MTT法检测各组细胞的增殖能力;流式细胞术检测细胞周期和凋亡情况。结果正常宫颈组织与宫颈癌组织间有25种蛋白的表达存在明显差异;体外实验结果显示,对照组与空质粒组survivin基因的转录与蛋白表达无明显变化,而pGCsi-sur组survivin基因转录和蛋白水平明显下降;MTT结果显示,pGCsi-sur组细胞增殖速度明显下降(P<0.01);流式细胞术检测显示,与对照组相比,pGCsi-sur组G1期细胞比例增加,G2期和S期细胞比例减少,并可检测到明显凋亡。结论宫颈癌组织与正常宫颈组织存在差异表达蛋白;survivin基因与宫颈癌的发生存在一定的相关性。
Objective Detect the differentially expressed proteins in cervical carcinoma to search for the relationship between surviving gene and carcinogenesis and development of cervical carcinoma.Methods With protein array technology we analyzed differentially expressed proteins in cervical carcinoma and normal cervical tissues;in vitro,the cervical cancer cell lines Hela were transfected with plasmids pGCsi-sur,to determine the expression levels of suvivin,the semi-quantitative RT-PCR and Western blot analysis with the samples extracted from treated and control cells were performed.The cells were also analyzed for cell cycle phase distribution and apoptosis rate by flow cytometry,and detected for the inhibition of cell proliferation by MTT.Results We found 25 kinds of proteins differentially expressed between cervical carcinoma and normal cervical tissues,some of them over-expression while others are down regulated in carcinoma.Survivin is over expressed in cervical cancer tissues.The results from semi-quantitative RT-PCR analysis and Western blot analyses for the Hela cell samples in pGCsi-sur group demonstrated that survivin siRNA could specifically reduce survivin expression.The results of MTT demonstrated that inhibition of cell proliferation were present in pGCsi-sur treated groups.Significant hypodiploidy peaks before G1 phase were present in treated cell samples in FCM analysis.Conclusion There are differentially expressed proteins in cervical carcinoma and normal cervical tissues,survivin may play roles the carcinogenesis and development of cervical carcinoma.
出处
《中国实验诊断学》
北大核心
2010年第10期1535-1538,共4页
Chinese Journal of Laboratory Diagnosis
