逆转录病毒介导的siRNA对宫颈癌细胞CXCR4表达的抑制及对侵袭能力的影响  

Down-Regulation of CXCR4 of Human Cervix Carcinoma Cells by Retrovirally Mediated RNA Interference

作　　者：康楷[1,2] 朱元方[3] 董晓静[3] 胡丽娜[3,4] 

机构地区：[1]重庆医科大学附属第一医院妇产科,重庆400016 [2]School of Health Sciences, University of Wollongong, Wollongong 2522, NSW, Australia [3]重庆医科大学附属第二医院妇产科,重庆400010 [4]四川大学华西医学中心第二医院妇产科,成都610041

出　　处：《中国生物工程杂志》2008年第7期87-91,共5页China Biotechnology

基　　金：国家自然科学基金资助项目(30371619)

摘　　要：目的:观察逆转录病毒介导RNA干扰抑制宫颈癌Caski细胞CXCR4基因表达的效率。方法:人工合成CXCR4特异性小干扰RNA(small interfering RNA,siRNA)片段,装入带有绿色荧光蛋白(GFP)的逆转录病毒载体pSOS,先转染PT67细胞包装成病毒,收获病毒上清,再将其转染Caski细胞,采用实时定量PCR和Western blotting观察CXCR4表达受抑的情况,并通过Transwells小室检测对肿瘤细胞侵袭能力的影响。结果:成功构建pSOS-CXCR4载体,并发现在24h,48h和72h,CXCR4mRNA的抑制率分别为29.9%,56.8%和62.8%,CXCR4蛋白的抑制率分别为43.6%,49.6%和62.9%。pSOS-CXCR4对Caski细胞的侵袭能力的抑制率为57.91%。结论:逆转录病毒介导RNA干扰能有效抑制宫颈癌细胞CXCR4表达并能降低癌细胞的侵润能力。Objective：To observe the effect of retrovirally mediated RNA interference targeting CXCR4 of human cervix carcinoma Caski cells. Methods ： Construct the fragment of CXCR4-siRNA, insert it into a retroviral vector which contain Green Flu Protein, named pSOS. Then, recombinated it by transfecting in PT67 cells, collect the supernatant of culture fluid, and use the supernatant transfect Caski cells. The expression of CXCR4 mRNA and protein were detected by Real-Time PCR and Western blotting. The abilities of invasion was detected by Matrigel invasion assay. Results： The retrovirus vectors pSOS-CXCR4 were constructed successfully. Compared with the negative control group, the inhibition rates of CXCR4 mRNA on the 24th, 48th and 72th hour were 29.9% , 56.8% and 62.8%, respectively. The inhibition rates of CXCR4 protein on the 24th, 48th and 72th hour were 43.6% ,49.6% and 62.9% , respectively. And the abilities of invasion was weaken when CXCR4＇s expression was inhibited, the inhibition rate was 57.91%. Conclusion： Retrovirally mediated RNA interference could effectively inhibit the expression of CXCR4 in human cervix carcinoma cells and weaken the the abilities of invasion.

关 键 词：RNA干扰 CXCR4 逆转录病毒 宫颈癌 

分 类 号：Q786[生物学—分子生物学]