摘要
目的研究MicroRNA-421(miR-421)促进前列腺癌DU145细胞增殖的作用及潜在的分子机制。方法培养前列腺癌DU145细胞,分为对照组、miR-阴性对照(NC)组、miR-421组、miR-421+pcDNA组、miR-421+pcDNA-细胞程序性死亡基因4(PDCD4),miR-NC组转染miR-NC、miR-421组转染miR-421、miR-421+pcDNA组转染miR-421及pcDNA质粒、miR-421+pcDNA-PDCD4组转染miR-421及pcDNA-PDCD4质粒,MTS法测定细胞增殖活力,荧光定量PCR测定PDCD4的mRNA表达水平,western blot测定PDCD4的蛋白表达水平,双荧光素酶报告基因实验验证miR-421与PDCD4的靶向结合。结果与对照组及miR-NC组比较,miR-421组的增殖活力增加,PDCD4的表达水平及野生型PDCD4双荧光素酶报告基因的荧光活力均降低(P<0.05);与miR-421+pcDNA组比较,miR-421+pcDNAPDCD4组的增殖活力降低,PDCD4的表达水平增加(P<0.05)。结论 miR-421对前列腺癌DU145细胞的增殖具有促进作用,靶向抑制PDCD4是miR-421发挥这一作用的潜在分子机制。
Objective To investigate the effect of microRNA-421(miR-421)on the proliferation of prostate cancer cell line DU145 and its potential molecular mechanism. Methods Prostate cancer DU145 cells were cultured and divided into control group,miR-negative control(NC)group,miR-421 group,miR-421+pcDNA group,miR-421+pcDNA-programmed cell death gene 4(PDCD4)group. MiR-NC group was transfected with miR-NC,miR-421 group was transfected with miR-421,miR-421 + pcDNA group was transfected with miR-421 and pcDNA plasmid,miR-421+pcDNA-PDCD4 group was transfected with miR-421 and pcDNA-PDCD4 plasmid,Cell proliferation activity was measured by MTS,PDCD4 mRNA expression was measured by FQ-PCR,PDCD4 protein expression was detected by Western blot,and the target binding of mir-421 and PDCD4 was verified by double luciferase reporter gene experiment. Results Compared with the control group and miR-NC group,the proliferation activity of the miR-421 group increased. The expression level of PDCD4 and the fluorescence activity of wild-type PDCD4 double luciferase reporter gene decreased(P<0.05). Compared with miR-421+pcDNA group,the proliferation activity of miR-421+pcDNA-PDCD4 group decreased,the expression level of PDCD4 increased(P<0.05). Conclusion miR-421 can promote the proliferation of prostate cancer cell line DU145 by targeting the inhibition of PDCD4 expression.
作者
李作为
郭航
邓超
燕东亮
田斌强
LI Zuowei;GUO Hang;DENG Chao;YAN Dongliang;TIAN Binqiang(Department of Urology,Shanghai University of Medicine&Health Sciences affiliated Sixth People's Hospital east,Shanghai,China,201306)
出处
《分子诊断与治疗杂志》
2020年第3期332-335,344,共5页
Journal of Molecular Diagnostics and Therapy
基金
上海健康医学院种子基金(HMSF-16-21-016)
上海健康医学院师资人才百人库
上海浦东新区发展基金(PKJ2014-Y07).
