miR-140抑制卵巢癌细胞增殖促进卵巢癌细胞凋亡及靶向抑制CMTM6表达  

Inhibition of proliferation, promotion of apoptosis, and target inhibition of CMTM6 expression by miR-140 in ovarian cancer cells

作　　者：杨俊 祝徳 段智慧 张子月 Yang Jun;Zhu De;Duan Zhihui;Zhang Ziyue(Bazhong Central Hospital Sichuan,Bazhong 636000;Department of Gerontology,General Hospital of Western Warzone,Sichuan 610036)

机构地区：[1]巴中市中心医院,巴中636000 [2]西部战区总医院老年科,四川610036

出　　处：《中国组织化学与细胞化学杂志》2022年第6期582-591,共10页Chinese Journal of Histochemistry and Cytochemistry

基　　金：2019年四川省卫生健康委员会科研课题19PJ070。

摘　　要：目的 探讨miR-140能否通过调控趋化素样因子超家族6(CMTM6)表达对卵巢癌细胞增殖、凋亡及程序性死亡配体1(PD-L1)表达产生影响。方法 将人源SKOV3卵巢癌细胞分为:对照组、inhibitor-NC组、miR-140 inhibitor组、mimic-NC组、miR-140 mimic组、miR-140mimic+pcDNA3.0-CMTM6组。MTT法、EdU染色和流式细胞仪、TUNEL染色分别检测细胞增殖和凋亡的发生情况。采用实时荧光定量PCR(qRT-PCR)检测CMTM6、PD-L1 mRNA表达水平。Western blot检测细胞CMTM6、PD-L1、增殖与凋亡相关蛋白表达水平。双荧光素酶报告基因实验验证CMTM6与miR-140的靶向关系。结果 与对照组相比,下调miR-140表达可增加SKOV3细胞存活率和EdU阳性率,上调c-Myc、cyclin D1、Bcl-2、CMTM6、PD-L1表达水平,抑制细胞凋亡和Bax、Caspase-3表达水平,而miR-140过表达可抑制SKOV3细胞存活率和EdU阳性率,下调c-Myc、cyclin D1、Bcl-2、CMTM6、PD-L1表达水平,促进细胞凋亡和Bax、Caspase-3表达;CMTM6过表达可逆转miR-140过表达对SKOV3细胞增殖的抑制、对凋亡的促进和对PD-L1表达的抑制。双荧光素酶报告基因实验结果显示,CMTM6是miR-140的靶基因。结论 miR-140可能通过下调CMTM6表达抑制卵巢癌细胞的增殖、促进卵巢癌细胞凋亡。Objective To investigate whether miR-140 could affect the proliferation, apoptosis and programmed death ligand 1(PD-L1) expression in ovarian cancer cells by regulating the expression of chemokine like factor superfamily 6(CMTM6). Methods Human SKOV3 ovarian cancer cells were divided into blank control group, inhibitor-NC group, miR-140 inhibitor group, mimic-NC group, miR-140 mimic group, and miR-140mimicn + pcDNA3.0-CMTM6 group. MTT method, EdU staining, flow cytometry and TUNEL staining were used to detect the occurrence of cell proliferation and apoptosis, respectively. Real-time fluorescence quantitative PCR(qRT-PCR) was used to detect the mRNA expression levels of CMTM6 and PD-L1. Western blot was used to detect the expression levels of CMTM6, PD-L1, proliferation and apoptosis related proteins. The dual luciferase reporter gene experiment verified the targeting relationship between CMTM6 and miR-140. Results Compared with the control group, down-regulation of miR-140 expression could promote SKOV3 cell survival and EdU positive rate, up-regulating expression levels of c-Myc, cyclin D1, Bcl-2, CMTM6, PDL1, inhibiting cell apoptosis, Bax, Caspase-3 expression levels, while overexpression of miR-140 can inhibit SKOV3 cell survival rate and EdU positive rate, down-regulating expression levels of c-Myc, cyclin D1, Bcl-2, CMTM6, PD-L1, and promoting cell apoptosis and levels of Bax and Caspase-3. Overexpression of CMTM6 could reverse the inhibition of proliferation and promotion of apoptosis of SKOV3 cells and expression inhibition of CMTM6 by overexpression of miR-140. The results of the dual luciferase reporter gene experiment showed that CMTM6 is the target gene of miR-140. Conclusion Overexpression of miR-140 may inhibit the proliferation and promote apoptosis of ovarian cancer cells by down-regulating the expression of CMTM6.

关 键 词：miR-140 趋化素样因子超家族6 卵巢癌 程序性死亡受体-配体1 

分 类 号：R737.31[医药卫生—肿瘤]