CXCR4-siRNA逆转录病毒载体对宫颈癌Caski细胞CXCR4基因表达的抑制作用

目的: 构建针对趋化因子受体CXCR4的RNA干扰逆转录病毒载体,检测由逆转录病毒介导的RNA干扰对人宫颈癌Caski细胞CXCR4基因表达的抑制作用. 方法: 人工合成CXCR4特异性小干扰RNA(small interfering RNA,siRNA)片段,将其插入带有绿色荧光蛋白(EGFP)的逆转录病毒载体质粒pSOS,测序正确后进行重组,转染PT67细胞包装成病毒,收获病毒上清用其转染Caski细胞,采用实时定量PCR和Western blot分别从mRNA水平和蛋白水平检测其对Caski细胞CXCR4表达的抑制作用. 结果: 成功构建pSOS-siCXCR4逆转录病毒载体,筛选出高效抑制序列,与对照组比较,在Caski细胞的mRNA水平,CXCR4-siRNA在24h、48h和72h对CXCR4 mRNA表达的抑制率分别为29.9%、56.8%和62.8%,而在蛋白水平对CXCR4蛋白的抑制率分别为43.6%、49.6%和62.9%. 结论: pSOS-HUS-siCXCR4干扰载体可以有效抑制Caski细胞中CXCR4表达.

Down-regulation of CXCR4 by Inducible Retrovirally Mediated RNA Interference Inhibits Human Cervix Carcinoma Cell Invasion in Vitro

Objective: It is suggested that the chemokine receptor CXCR4 plays an important role in the invasion and metastasis of cervical cancer, but there are few published articles about down-regulating the CXCR4 expression to prevent the metastasis of uterine cervix cancer. The aim of this study was to construct RNA inter- fering retrovirus vector of CXCR4 and to investigate the expression of CXCR4 receptor in human cervix carci- noma Caski cells interfered with retrovirally mediated RNA interference targeting CXC...