摘要
目的探究微小RNA378(miR-378)与c-Myc表达的关系及其与卵巢癌细胞增殖能力的影响。方法培养对数生长期人卵巢癌细胞系SKOV3,慢病毒转染法外源性上调SKOV3细胞中miR-378的表达作为实验组,同时转染空白载体作为对照组,Taqman实时荧光定量PCR(RT-q PCR)技术检测两组细胞miR-378及c-Myc mRNA表达水平,免疫印迹实验(Western Bolt)检测两组细胞c-Myc蛋白表达水平,CCK-8实验检测两组细胞增殖能力的情况,克隆形成实验检测两组细胞克隆形成能力的情况。结果实验组miR-378的表达水平显著高于对照组细胞(P<0.001);实验组c-Myc mRNA表达水平显著低于对照组细胞(P<0.05);实验组c-Myc蛋白表达水平同样显著低于对照组细胞(P<0.05);实验组细胞增殖能力在48及72h时显著低于对照组细胞(P<0.05);实验组细胞克隆形成能力显著低于对照组细胞(P<0.01)。结论 miR-378可能通过抑制原癌基因c-Myc的表达,介导卵巢癌细胞增殖能力的下调。
Objective To explore the relationship between microRNA378（ miR-378） expression and c-Myc expression and the effects on proliferation ability of ovarian cancer cells. Methods Human ovarian cancer cell line SKOV3 in logarithmic growth phase was cultured,lentiviral transfection method was used to transfect exogenous up-regulated SKOV3 cells（ experimental group）,SKOV3 cells transfected by empty vector were selected as control group. Taqman real-time fluorescence quantitative PCR（ RT-q PCR） was used to detect the expression levels of miR-378 and c-Myc mRNA in the two groups. Western blot was used to detect the expression levels of c-Myc protein in the two groups. CCK-8 assay was used to detect cell proliferation abilities in the two groups. Clone formation assay was used to detect clone formation abilities in the two groups. Results The expression level of miR-378 in experimental group was statistically significantly higher than that in control group（ P〈0. 001）. The expression level of c-Myc mRNA in experimental group was statistically significantly lower than that in control group（ P〈0. 05）. The expression level of c-Myc protein in experimental group was statistically significantly lower than that in control group（ P〈0. 05）. Cell proliferation abilities at 48 and 72 hours in experimental group were statistically significantly lower than those in control group（ P〈0. 05）. Cell clone formation ability in experimental group was statistically significantly lower than that in control group（ P〈0. 01）. Conclusion miR-378 may downregulate the proliferation ability of ovarian cancer cells by inhibiting the expression of oncogene c-Myc.
作者
陈小平
任新萍
杜依蓓
薛金玲
CHEN Xiao-Ping;REN Xin-Ping;DU Yi-Bei;et al.(Department of Gynecology and Obstetrics, the First People＇s Hospital of Yancheng , Yancheng , Jiangsu 224000, Chin)
出处
《中国妇幼保健》
CAS
2018年第11期2562-2565,共4页
Maternal and Child Health Care of China
基金
江苏省333工程资助项目(BRA2016218)
