间皮素基因对人卵巢癌细胞株SKOV3生长和迁移的影响  

Effect of mesothelin gene on the growth and migration of human ovarian cancer cell line SKOV3

作　　者：赵楠[1] 刘晓芳 杨雪[1] 张俐佳 刘岩[1] ZHAO Nan;LIU Xiao-fang;YANG Xue;ZHANG Li-jia;LIU Yan(Office of the National Traditional Chinese Medicine Clinical Research Base,First Affiliated Hospital of Heilongjiang University of Traditional Chinese Medicine;Department of Oncology,Heilongjiang University of Traditional Chinese Medicine,Harbin 150040,China)

机构地区：[1]黑龙江中医药大学附属第一医院国家中医临床研究基地项目办公室 [2]黑龙江中医药大学针灸推拿学院肿瘤科,黑龙江哈尔滨150040

出　　处：《哈尔滨医科大学学报》2020年第6期589-593,共5页Journal of Harbin Medical University

基　　金：黑龙江省博士后资助项目(LBH-Z16201);黑龙江省教育厅优秀创新人才项目(UNPYSCT-2018225);黑龙江省中医药科研项目(ZHY2020-111)。

摘　　要：目的探讨间皮素(mesothelin,MSLN)基因干扰对人卵巢癌细胞株SKOV3生长、迁移的影响。方法培养人卵巢癌SKOV3细胞,分别设置空白对照组、基因敲低组、阴性对照组。基因敲低组转染MSLN小干扰核糖核酸(siRNA),阴性对照组转染乱序非特异性siRNA,转染48 h空白对照组不进行细胞转染。收集细胞,通过实时荧光定量聚合酶链式反应(RT-qPCR)和蛋白免疫印迹(WB)检测MSLN信使核糖核酸(mRNA)和蛋白相对表达量;通过四唑盐(MTT)法检测细胞增殖能力;通过细胞划痕实验检测细胞迁移能力;通过RT-qPCR和WB检测细胞中癌抗原(CA125)和基质金属蛋白酶9(MMP-9)mRNA和蛋白相对表达量。结果 MSLN mRNA和蛋白表达水平基因敲低组显著低于空白对照组与阴性对照组(P<0.05);基因敲低组细胞增殖抑制率显著高于空白对照组与阴性对照组(P<0.05);基因敲低组细胞迁移率小于空白对照组与阴性对照组(P<0.05);CA125、MMP-9 mRNA和蛋白表达水平基因敲低组细胞均低于空白对照组与阴性对照组(P<0.05)。结论通过siRNA干扰对MSLN基因敲低可抑制人卵巢癌细胞株SKOV3的增殖和迁移能力,推测其机制可能与CA125、MMP-9 mRNA和蛋白表达水平下调有关。Objective To explore the effect of mesothelin(MSLN) gene interference on the growth and migration of human ovarian cancer cell line SKOV3. Methods Human ovarian cancer SKOV3 cells were cultured. Then cells were divided into 3 groups with 3 replicates in each group, which were blank control group, knockdown group and negative control group. The knockdown group was transfected with MSLN siRNA by liposome transfection technique, the negative control group was transfected with disordered non-specific siRNA, and the blank control group was not performed. Cells were transfected for 48 h. Cells were collected, and MSLN mRNA and protein expression levels were detected by real-time quantitative PCR(RT-qPCR) and Western blot(WB). Cell proliferation inhibition rates was detected by methyl thagolyl tetragoliam(MTT) method. Cell migration ability by cell scratch test was detected. Relative expression levels of CA125 and MMP-9 mRNA and protein in cells were detected by RT-qPCR and WB. Results The expression levels of MSLN mRNA and protein in the knockdown group was significantly lower than the blank control group and the negative control group(P<0.05). The inhibition rate of cell proliferation in knockdown group was lower than the blank control group and the negative control group(P<0.05). The cell migration rate in the knockout group was lower than blank control group and the negative control group(P<0.05). The expression levels of CA125 and MMP-9 mRNA and protein in the knockdown group were lower than the blank control group and the negative control group(P<0.05). Conclusion Knocking down the MSLN gene by siRNA interference can inhibit the proliferation and migration ability of human ovarian cancer cell line SKOV3, suggesting that the mechanism may be related to the down-regulation of CA125, MMP-9 mRNA and protein expression levels.

关 键 词：间皮素 小干扰RNA 人卵巢癌细胞 增殖 迁移 

分 类 号：R737.31[医药卫生—肿瘤]