RBM8A基因对子宫内膜癌HEC-1A细胞增殖、迁移和凋亡的作用及其机制  

Effect and Mechanism of RBM8A on Proliferation,Migration and Apoptosis of Human Endometrial Cancer HEC-1A Cells

作　　者：谭冬梅 张静静[2] 师一民 韩赛[2] 耿炜 孙建一 王雅玉 张秀荣 TAN Dongmei;ZHANG Jingjing;SHI Yimin;HAN Sai;GENG Wei;SUN Jianyi;WANG Yayu;ZHANG Xiurong(TCM Gynecology,Shandong Provincial Maternal and Child Health Care Hospital,Ji’nan 250000,China;Department of Obstetrics and Gynecology,Qilu Hospital of Shandong University,Ji’nan 250000,China)

机构地区：[1]山东省妇幼保健院中医妇科,济南250000 [2]山东大学齐鲁医院妇产科,济南250000

出　　处：《肿瘤防治研究》2023年第1期27-32,共6页Cancer Research on Prevention and Treatment

基　　金：山东省中医药科技项目(2021M202)。

摘　　要：目的探讨沉默RBM8A基因表达对子宫内膜癌HEC-1A细胞生物学行为的影响及其可能机制。方法设计RBM8A基因靶向的发夹状shRNA,筛选出最佳shRNA沉默片段,构建携带有目的基因的重组慢病毒干扰载体并感染HEC-1A细胞,通过嘌呤霉素筛选出稳定敲低RBM8A基因的细胞作为实验组(shRBM8A),同时设计无意义序列的shRNA作为对照组(shControl)。CCK-8法检测细胞增殖能力,Transwell检测细胞迁移和侵袭能力,流式细胞计数法检测细胞凋亡,Western blot法分析凋亡相关蛋白、上皮间质转化(EMT)信号转导通路相关蛋白的表达。结果与shControl组比较,敲低RBM8A后HEC-1A细胞增殖减慢,凋亡率增加,且迁移与侵袭能力受到明显抑制(P<0.05),凋亡相关蛋白cleaved-caspase 9和cleaved-caspase 3表达增加,EMT相关蛋白E-cadherin表达升高,Vimentin表达降低。结论沉默RBM8A基因可抑制子宫内膜癌细胞增殖、迁移、侵袭并促进细胞凋亡,抑制EMT信号转导通路可能是其作用机制。Objective To investigate the effect of silenced RBM8A gene on the biological behavior(proliferation,migration,and apoptosis)of human endometrial cancer HEC-1A cells and its possible mechanism.Methods The hairpin shRNA targeted by the RBM8A gene was designed,and the best shRNA silencing fragment was screened.The recombinant lentiviral interference vector carrying the target gene was constructed and used to infect HEC-1A cells.Cells with stable knockdown of RBM8A gene were screened by puromycin as the experimental group(shRBM8A),while the shRNA of nonsense sequence was designed as the control group(shControl).CCK-8 method was used to detect cell proliferation,and flow cytometry was used to detect cell apoptosis.Transwell assay was used to detect cell migration and invasion.Western blot was used to analyze the expression of apoptosis-related proteins and EMT signal transduction pathway related proteins.Results In comparison with the shControl group,after RBM8A knockdown,HEC-1A cell proliferation was reduced,apoptosis was increased,migration and invasion ability were significantly inhibited(P<0.05),the expression of apoptosis-related proteins cleaved caspase 9 and caspase 3 increased,EMTrelated protein E-cadherin expression increased,and Vimentin expression decreased.Conclusion RBM8A gene silencing can inhibit the proliferation,migration,and invasion and promote the apoptosis of endometrial cancer cells.The inhibition of EMT signal transduction pathway may be its mechanism.

关 键 词：子宫内膜癌 RBM8A 增殖 迁移 凋亡 上皮间质转化 

分 类 号：R737.33[医药卫生—肿瘤]