RNA干扰沉默宫颈鳞癌细胞中HIF-1a基因对survivin基因表达的影响

摘要

目的：研究 RNA 干扰对宫颈鳞癌 SiHa 细胞中 HIF-1α及survivin 的抑制效应。方法使用不同缺氧时间分别缺氧 SiHa 细胞，以Westernblot 法检测细胞中 HIF-1a 蛋白的表达变化，选择可以引起HIF-1a 蛋白表达增加的缺氧培养时间。进行缺氧后用 Westernblot 法和逆转录聚合酶链反应检测 HIF-1α及 survivin 的表达。结果 SiHa细胞 HIF-1α的表达在缺氧处理时较常氧培养时增加，在缺氧后24～48小时内明显升高，60小时略有降低，72～96 h 逐渐减少。氯化钴培养后 SiHa 细胞的 HIF-1a 蛋白及 survivin 蛋白表达增加，并与氯化钴浓度有关。通过 RNA 干扰技术抑制 HIF-1a 基因的干扰后的细胞中 HIF-1a 及 survivin 表达同时减少。结论在 SiHa 细胞中，HIF-1α基因和 survivin 表达具有高度的相关性，并且抑制 HIF-1α表达可能通过抑制 survivin 而促进癌细胞的凋亡。%Objective To explore the inhibition effect of RNA interference to HIF-1 alpha and survivin in cervical squamous carcinoma SiHa cells. Methods Deprivate oxygen to SiHa cells in different time, then detecte the changing of HIF-1a protein by Westernblot and choosing the right time which can increase the expression of HIF-1a. Use Westernblot and RT-PCR to detect the expression of HIF - 1 alpha and survivin. Results The HIF-1a expression in SiHa cells was increased by oxygen deprivation, and rised in 24~48 h, decreased a little in 60 h and much more in 72~96 h. The expression of HIF-1a and survivin is related to the concentration of CoCl2, and was decreased by using RNA interference. Conclusion In SiHa cells, the expression of HIF-1a is highly related to survivin so that suppression of HIF-1a could promote apoptosis of cancer cells.