抑制miR-630表达对宫颈癌细胞紫杉醇敏感性的影响及机制  

Effect and mechanism of inhibiting microRNA-630 expression on sensitivity of cervical cancer cells to paclitaxel

作　　者：丁鑫[1] 王乾印 马亮亮 DING Xin;WANG Qianyin;MA Liangliang(Qinghai Red Cross Hospital,Xining 810000,China)

机构地区：[1]青海红十字医院,西宁810000

出　　处：《山东医药》2018年第42期9-12,共4页Shandong Medical Journal

基　　金：青海省科技计划项目(2015-ZJ-772)

摘　　要：目的观察制微小RAN-630(miR-630)表达对宫颈癌细胞紫杉醇(PTX)敏感性的影响,并探讨其机制。方法将宫颈癌PTX耐药细胞株He La-PTX分为3组,抑制组和空载组分别转染miR-630小干扰RNA质粒、对照空载质粒30 nmol/L,空白对照组不转染,48 h后收集细胞。采用MTT法检测PTX对He La-PTX细胞的半数抑制浓度(IC50),实时荧光定量PCR法检测细胞中的miR-630、凋亡蛋白酶活化因子(APAF-1) mRNA,流式细胞术检测细胞的凋亡情况,Western blotting法检测凋亡相关蛋白聚腺苷二磷酸核糖聚合酶(PARP)、Caspase1、Caspase3。结果抑制组、空载组、空白对照组PTX IC50分别为(35. 61±2. 87)、(142. 57±3. 63)、(144. 31±2. 42) nmol/L,抑制组PTX IC50低于NC组和空白对照组(P均=0. 000)。与空载组及空白对照组比较,抑制组miR-630表达量降低而APAF-1 mRNA表达量增加(P均=0. 000)。抑制组、空载组、空白对照组细胞凋亡率分别为19. 60%±2. 56%、10. 20%±1. 08%、8. 00%±1. 53%,抑制组细胞凋亡率高于空载组和空白对照组(P均=0. 000)。与空载组及空白对照组比较,抑制组PARP、Caspase1、Caspase3相对表达量增加(P均=0. 000)。结论抑制miR-630表达能增强宫颈癌细胞对PTX的敏感性,这可能是通过调节细胞凋亡及APAF-1基因表达实现的。Objective To observe the effect of inhibiting microRNA-630 expression on sensitivity of cervical cancer cells to paclitaxel(PTX),and to explore the possible mechanism.Methods The PTX resistant cell line HeLa-PTX of cervical cancer was divided into three groups:the inhibition group,unloaded group,and blank control group.The small interfering RNA plasmid MiR-630 and control of unloaded plasmid 30 nmol/L were transfected into the cells of the inhibition group and the unloaded group,respectively,and the cells in the blank control group were not transfected.We collected the cells at 48 h,and MTT assay was used to determine the half inhibitory concentration of PTX on the HeLa-PTX cell(IC 50),real-time fluorescent quantitative PCR was used to detect the miR-630 and apoptotic protease activating factor(APAF-1)mRNA,and flow cytometry was used to detect the apoptosis,and Western blotting was applied to detect the expression levels of poly adp-ribose polymerase(PARP,Caspase1,and Caspase3).Results In the inhibition group,unloaded group,and blank control group,PTX IC 50 was(35.61±2.87),(142.57±3.63),and(144.31±2.42)nmol/L,respectively.The PTX IC 50 in the inhibition group was lower than that in the unloaded group and the blank control group(P=0.000).Compared with the unloaded group and blank control group,the miR-630 expression decreased and the APAF-1 mRNA expression increased in the inhibition group(P=0.000),the apoptosis rates in the inhibition group,the unloaded group and the blank control group were 19.60%±2.56%,10.20%±1.08%and 8%±1.53%,respectively,and the apoptosis rate of the inhibition group was higher than that of the unloaded group and the blank control group(P=0.000).Compared with the unloaded group and blank control group,the relative expression levels of PARP,Caspase1 and Caspase3 increased in the inhibition group(P=0.000).Conclusion Inhibiting microRNA-630 expression can enhance the sensitivity of cervical cancer cells to PTX possibly by regulating apoptosis and APAF-1 gene expression.

关 键 词：宫颈癌 微小RAN-630 紫杉醇 细胞耐药 细胞凋亡 凋亡蛋白酶活化因子 聚腺苷二磷酸核糖聚合酶 含半胱氨酸的天冬氨酸蛋白水解酶 

分 类 号：R737.33[医药卫生—肿瘤]