F10基因siRNA干扰对宫颈癌细胞侵袭和凋亡的影响及机制研究  

Effect of silencing F10 gene by RNA interference on invasion and apoptosis of cervical carcinoma cells and molecular mechanism

作　　者：韦娟冰[1] 林静 WEI Juan-Bing;LIN Jing(Gynecology Department,the First Affiliated Hospital of Fujian Medical University,Fuzhou 350000,China)

机构地区：[1]福建医科大学附属第一医院妇产科,福州350000 [2]福建医科大学基础医学院组胚教研室,福州350000

出　　处：《中国免疫学杂志》2019年第13期1584-1589,共6页Chinese Journal of Immunology

基　　金：福建省科技计划项目(No.2015Y6001)

摘　　要：目的:以小干扰RNA(siRNA)抑制人宫颈癌Hela细胞中F10基因的表达,研究下调F10基因的表达对细胞侵袭及凋亡的影响,并对其作用机制进行初步探讨。方法:采用荧光定量聚合酶链式反应(qRT-PCR)和蛋白印迹法(Western blot)检测宫颈癌细胞株Hela及正常宫颈上皮细胞株H8中F10 mRNA和蛋白表达水平。在宫颈癌Hela细胞中转染F10 siRNA,并以转染siRNA control的细胞为阴性对照,以不做任何处理的细胞为空白对照组,转染后24 h,分别以qRT-PCR和Western blot检测转染效率。Transwell实验检测细胞侵袭能力,流式细胞术检测细胞凋亡情况,Western blot检测细胞中基质金属蛋白酶-2(MMP-2)、剪切的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved Caspase-3)和磷酸化的信号转导与转录因子3(p-STAT3)蛋白表达水平。结果:在宫颈癌细胞中F10 mRNA和蛋白呈高表达。转染F10 siRNA能够抑制Hela细胞中F10 mRNA和蛋白的表达。下调F10的表达能够明显抑制Hela细胞的侵袭能力,诱导细胞凋亡,下调MMP-2和p-STAT3的表达,上调细胞中Cleaved Caspase-3的表达。结论: F10在宫颈癌细胞中呈高表达,抑制其表达能够抑制Hela细胞的侵袭能力,诱导细胞发生凋亡,其作用机制可能与下调MMP-2和p-STAT3的表达,激活Caspase-3有关。Objective: To study the effect of down-regulation of F10 gene expression on cell invasion and apoptosis by inhibiting the expression of F10 gene in human cervical cancer Hela cells with small interfering RNA(siRNA),and to explore its mechanism. Methods: qRT-PCR and Western blot were used to detect the expression of F10 in Hela cervical cancer cell line and H8 cervical epithelial cell line.F10 siRNA was transfected into Hela cells of cervical cancer,and the cells transfected with siRNA control were used as negative control.The cells without any treatment were used as blank control group.The transfection efficiency was detected by qRT-PCR and Western blot 24 hours after transfection.Transwell assay was used to detect cell invasion ability,and apoptosis was detected by flow cytometry.Western blot was used to detect the expression levels of MMP-2,Cleaved Caspase-3 and p-STAT3 in cells. Results: F10 mRNA and protein were highly expressed in cervical cancer cells.Transfection of F10 siRNA inhibited the expression of F10 mRNA and protein in Hela cells.Down-regulation of F10 expression could significantly inhibit the invasion ability of Hela cells,induce apoptosis,down-regulate the expression of MMP-2 and p-STAT3,and up-regulate the expression of Cleaved Caspase-3 in cells. Conclusion: F10 is highly expressed in cervical cancer cells.Inhibition of its expression can inhibit the invasion of Hela cells and induce apoptosis.The mechanism may be related to down-regulation of MMP-2 and p-STAT3 expression and activation of Caspase-3.

关 键 词：F10基因 宫颈癌HELA细胞 侵袭 凋亡 

分 类 号：R737.33[医药卫生—肿瘤]