宫颈癌细胞C33a下调Raptor表达后对增殖及化疗敏感性的研究  

Study on the effect of down- regulation of Raptor expression in cervical cancer C33a cells on proliferation and chemosensitivity

作　　者：秦宇[1] 赵雪娇[1] 王婷[1] 朱涛[1] 

机构地区：[1]华中科技大学附属同济医院妇产科,湖北武汉430030

出　　处：《中国妇幼保健》2014年第9期1411-1413,共3页Maternal and Child Health Care of China

基　　金：国家自然基金项目〔30801340〕

摘　　要：目的:探讨通过反义RNA技术下调Raptor基因的表达,研究宫颈癌细胞株C33a中Raptor下调后宫颈癌细胞增殖及对化疗药物的敏感性。方法:C33a细胞分3组,实验组和阴性对照组分别转染Raptor siRNA和阴性对照siRNA,空白对照组不做任何处理。通过荧光实时定量PCR和蛋白印记法分别检测C33a转染siRNA后Raptor在mRNA和蛋白水平的表达,CCK-8法检测各组细胞的增殖和对顺铂敏感性的变化,流式细胞仪检测各组及加入顺铂后的凋亡情况。结果:转染siRNA 72 h后,Raptor分别在mRNA水平及蛋白水平较阴性对照下调为82%及78%(P<0.05),转染72 h后CCK-8法测细胞增殖,A值由空白对照组1.828±0.059、阴性对照组1.723±0.044明显降低为Raptor siRNA组的1.349±0.040(P<0.05),各组加入8μM顺铂后,Raptor siRNA组凋亡率为(40.14±1.57)%较空白对照组(21.76±1.23)%和阴性对照组(20.81±1.45)%明显增加(P<0.05)。结论:Raptor siRNA能显著下调C33a细胞中Raptor在mRNA和蛋白水平的表达,抑制C33a细胞的增殖,增加对化疗药物顺铂的敏感性和促凋亡作用。Objective： To study proliferation and chemotherapy of cervical cancer C33a cells after clown - regulation of Raptor by antisense RNA technique. Methods： Cervical cancer C33a cells were divided into three groups, the cells in experimental group and negative control group were transfeeted with Raptor siRNA and negative control siRNA, respectively; the cells in blank control group had no treatment; florescent quantitative RT - PCR and western blotting were used to detect the expressions of Raptor mRNA and protein after transfection with siRNA, CCK - 8 method was used to detect proliferations and changes of chemosensitivity to cisplatin, flow cytometry was used to measure apoptosis. Results： After transfection for 72 hours, the expressions of Raptor mRNA and protein in experimental group decreased by 82% and 78%, respectively, compared with negative control group, there were statistically significant differences （P 〈 0. 05）, A values in blank control group, negative control group, and experimental group were （ 1. 828 ±0. 059）, （ 1. 723± 0. 044）, and （ 1. 349 ± 0. 040）, respectively （P 〈0. 05） ; the apoptotic rate in experimental group was （40. 14 ±1.57） %, which were higher than those in blank control group [ （21.76± 1.23） % ] and negative control group [ （20. 81 ± 1.45） % ] （P 〈 0. 05） . Conclusion ： Raptor siRNA can downregulate the expressions of Raptor mRNA and protein, inhibit proliferation, improve chemosensitivity and promote apoptosis in cervical cancer C33a ceils.

关 键 词：宫颈癌 C33a细胞 RAPTOR RNA干扰 凋亡 

分 类 号：R737.33[医药卫生—肿瘤]