HPV16E7靶向小干扰RNA对SiHa细胞增殖凋亡及6种抑癌基因的影响  

Effects of a small interfering RNA targeting HPV16E7 on proliferation and apoptosis of SiHa cells and expressions of six tumor suppressor genes

作　　者：龙嘉[1] 李黎明[2] 许翠[2] 杨佳[2] 蒋明军[2] 

机构地区：[1]深圳大学第一附属医院,深圳市第二人民医院皮肤科,518035 [2]中国医学科学院 北京协和医学院 皮肤病研究所 江苏省皮肤病与性病分子生物学重点实验室

出　　处：《中华皮肤科杂志》2016年第10期717-721,共5页Chinese Journal of Dermatology

基　　金：国家自然科学基金（31470274）;江苏省临床医学研究中心项目（BL2012003）;中央级公益性科研院所基本科研业务费（2016RC310025）

摘　　要：目的：探讨HPV16E7在SiHa细胞株中对抑癌基因（MT1G、NMES1、RRAD、SFRP1、SPARC、TFPI2）的表达及细胞增殖能力和凋亡水平的影响。方法 SiHa细胞转染E7SiRNA 48 h后，qPCR检测E7及6种抑癌基因mRNA水平变化，CCK-8检测细胞增殖能力改变，流式细胞仪检测细胞凋亡水平。结果 SiHa细胞转染后48 h，qPCR结果显示实验组E7 mRNA显著降低（0.25±0.036，P〈0.05）；6种抑癌基因的mRNA水平均显著增加（MT1G 1.403±0.190、NMES11.720±0.060、RRAD 1.390±0.160、SFRP11.493±0.120、SPARC 2.157±0.144、TFPI22.060±0.122，P〈0.05）。细胞增殖结果显示，与阴性对照组及空白组比较，实验组细胞增殖能力显著降低（0.554±0.130，P〈0.05），阴性对照组和空白组之间差异无统计学意义（P〉0.05）。细胞凋亡结果提示，实验组细胞凋亡细胞频率为9.222%较阴性对照组（0.246%）及空白组（0.123%）明显增加（P〈0.05），空白组与阴性对照组差异无统计学意义（P〉0.05）。结论 HPV16导致细胞恶性转化过程中，E7可能通过抑制（MT1G、NMES1、RRAD、SFRP1、SPAR、TFPI2）6种抑癌基因的表达参与作用。Objective To evaluate effects of human papilloma virus（HPV）16E7 on expressions of six tumor suppressor genes（including MT1G, NMES1, RRAD, SFRP1, SPARC and TFPI2）in a cell line SiHa, as well as on its proliferation and apoptosis. Methods SiHa cells were divided into two groups to be transfected with a small interfering RNA targeting HPV16E7（E7SiRNA, experimental group）and an empty vehicle（negative control group） respectively, with SiHa cells receiving no treatment serving as the blank control group. After 48 hours, qPCR was performed to measure the mRNA expressions of E7 and six tumor suppressor genes, CCK-8 assay to evaluate cellular proliferative activity, and flow cytometry to assess apoptosis of SiHa cells. Results At 48 hours after the transfection, the experimental group showed significantly decreased E7 mRNA expression（0.25 ± 0.036, P〈0.05）, but increased mRNA expressions of the six genes（MT1G 1.403 ± 0.190, NMES1 1.720 ± 0.060, RRAD 1.390 ± 0.160, SFRP1 1.493 ± 0.120, SPARC 2.157 ± 0.144, TFPI2 2.060 ± 0.122, all P 〈 0.05）. The proliferative activity of SiHa cells was significantly decreased（0.554 ± 0.130 vs. 1.028 ± 0.236 and 1.220 ± 0.126, both P〈0.05）, but the apoptosis rate was significantly increased（9.222%vs. 0.246%and 0.123%, both P〈0.05）in the experimental group compared with the negative control group and blank control group. No significant differences were observed between the negative control group and blank control group in proliferative activity or apoptosis rate of SiHa cells（both P〉0.05）. Conclusion E7 may participate in HPV16-induced cellular malignant transformation by suppressing the mRNA expressions of 6 tumor suppressor genes, including MT1G, NMES1, RRAD, SFRP1, SPAR and TFPI2.

关 键 词：人乳头瘤病毒16 乳头瘤病毒E7蛋白质类 细胞增殖 细胞凋亡 SiRNA 抑癌基因 

分 类 号：R730.2[医药卫生—肿瘤]