siRNA抑制人卵巢癌SKOV-3细胞MDM2的表达及细胞增殖的研究  

siRNA inhibits MDM2 expression and cell proliferation on ovarian cancer SKOV-3 cells

作　　者：王莹 贾志勇 赵燕颖[3] WANG Ying;JIA Zhi-yong;ZHAO Yan-ying(Department of Obstetrics and Gynecology,The Baogang No.3 Hospital of Hongci Medical Group,Baotou 014010,China)

机构地区：[1]内蒙古弘慈医疗集团包钢三医院妇产科,内蒙古包头014010 [2]内蒙古科技大学包头医学院第一附属医院普外二科 [3]吉林大学第四医院

出　　处：《中国实验诊断学》2021年第3期421-425,共5页Chinese Journal of Laboratory Diagnosis

摘　　要：目的研究siRNA对人卵巢癌细胞SKOV-3细胞MDM2基因表达的抑制作用及对肿瘤细胞增殖和凋亡的影响。方法体外合成2对针对MDM2基因的siRNA,经脂质体转染导入SKOV-3细胞;转染siRNA MDM2(简称si-MDM2)转染阴性对照质粒到SKOV-3,并设只加转染试剂作空白对照组,采用MTT法检测si-MDM2对细胞增殖的抑制作用。流式细胞仪检测细胞凋亡,并用Real-time PCR和Western blot分别从mRNA水平和蛋白水平检测si-MDM2对MDM2的抑制作用。结果 MTT结果表明转染siRNA MDM2后细胞生长受到抑制,与对照组比差异具有显著性(P<0.05);同时细胞发生凋亡和细胞周期阻滞,转染siRNA MDM2-1,2的凋亡率分别为47.8%和39.2%,转染阴性对照质粒和对照组未检测出明显的增殖抑制和凋亡改变。转染siRNA MDM2的细胞的MDM2基因表达分别下调到对照组的48.41%和46.51%;转染阴性对照质粒的MDM2基因没有明显改变。转染siRNA MDM2的细胞的MDM2蛋白表达水平同样显著低于对照组细胞(P<0.05)。结论 siRNA MDM2可以有效抑制SKOV-3细胞株中MDM2的表达,并抑制细胞增殖,诱导细胞凋亡。Objective To study the inhibitory effects of siRNA on MDM2 expression on ovarian cancer SKOV-3 cells and cell proliferation.Methods siRNA targeting MDM2(PGCsilencer TM-MDM2-siRNA)was constructed and transfercted into SKOV-3 cells.Meanwhile,control siRNA was transfected and the transfection agent was used as the comparison.Cell proliferation was measured with MTT assay.Then the interference effect was detected by real-time PCR and Western blot method.Results Real-time PCR result showed that MDM2 mRNA expression levels were reduced to 48.41% and 46.51% after being transfected with siMDM2-1 and siMDM2-2;The difference between control siRNA and liposome group was not obvious(P>0.05).The expression level of protein in siRNA MDM2 experimental group was statistically significantly lower than that in control group(P<0.05).The results of MTT and flow cytometer showed that siRNA could suppress the proliferation and induce apoptosis of SKOV-3 cells.Conclusion siRNA can effectively down-regulate MDM2 expression in SKOV-3 cells line,inhibit the cell proliferation and induce apoptosis.

关 键 词：SIRNA 卵巢癌细胞 MDM2 Western blot 

分 类 号：R737.31[医药卫生—肿瘤]