微RNA-128-3p靶向SLC39A7基因促进卵巢癌细胞内质网应激诱导的细胞凋亡  

miR-128-3p promotes endoplasmic reticulum stress-induced apoptosis in ovarian cancer cells by targeting SLC39A7

作　　者：李静萍[1] 冯勤梅[1] 杨芳[1] 郭亚丽[1] Li Jingping;Feng Qinmei;Yang Fang;Guo Yali(Department of Gynecology,Shanxi Provincial People’s Hospital,Taiyuan,Shanxi 030002,China)

机构地区：[1]山西省人民医院妇科,太原030002

出　　处：《中华生物医学工程杂志》2019年第5期558-564,共7页Chinese Journal of Biomedical Engineering

基　　金：山西省自然科学基金(201701D121156)。

摘　　要：目的探讨微RNA(miR)-128-3p/SLC39A7分子轴在卵巢癌细胞内质网应激诱导的细胞凋亡中的作用。方法应用3μmol/L的衣霉素(TM)诱导OVCAR-3细胞内质网应激,采用实时荧光定量PCR(qRT-PCR)和蛋白免疫印迹(Western blotting)检测未经TM处理及TM处理24 h后人卵巢癌细胞OVCAR-3内miR-128-3p和SLC39A7表达水平差异。构建过表达miR-128-3p或敲减SLC39A7的OVCAR-3细胞株,流式细胞术检测过表达miR-128-3p或敲减SLC39A7对内质网应激状态下的OVCAR-3细胞凋亡的影响,Western blotting检测SLC39A7蛋白、内质网应激蛋白ATF4和CHOP及凋亡蛋白Cleaved-caspase-3的表达。双荧光素酶报告基因实验和Western blotting验证miR-128-3p和SLC39A7的靶向关系。结果TM处理可显著促进OVCAR-3细胞miR-128-3p的表达,抑制SLC39A7的表达。过表达miR-128-3p或敲减SLC39A7均可促进内质网应激状态下的卵巢癌细胞凋亡。SLC39A7是miR-128-3p的靶基因,miR-128-3p可负性调控SLC39A7的表达。过表达SLC39A7可部分逆转miR-128-3p对内质网应激状态下的卵巢癌细胞凋亡的影响。结论miR-128-3p通过靶向下调SLC39A7表达促进内质网应激诱导的卵巢癌细胞凋亡。Objective To investigate the role of miR-128-3p/SLC39A7 molecular axis in apoptosis of ovarian cancer cells induced by endoplasmic reticulum stress.Methods 3μmol/L tunicamycin(TM)was used to induce endoplasmic reticulum stress in OVCAR-3 cells.The expression levels of miR-128-3p and SLC39A7 in untreated ovarian cancer OVCAR-3 cells and those treated with TM for 24 h were detected by qRT-PCR and Western blotting.OVCAR-3 cell lines over-expressing miR-128-3p or knocked down of SLC39A7 were constructed.Flow cytometry was used to detect the effect of over-expressing miR-128-3p or SLC39A7 knockdown on apoptosis of OVCAR-3 cells with endoplasmic reticulum stress.The expression levels of SLC39A7,endoplasmic reticulum stress-related proteins ATF4 and CHOP,and apoptosis-related protein Cleaved-caspase-3 were detected by Western blotting.Dual luciferase reporter gene assay and Western blotting were used to verify the targeting relationship between miR-128-3p and SLC39A7.Results The treatment with TM significantly promoted the expression of miR-128-3p but inhibited the expression of SLC39A7 in OVCAR-3 cells.Over-expressing miR-128-3p or knocking down SLC39A7 promoted the apoptosis of ovarian cancer cells with endoplasmic reticulum stress.SLC39A7 was identified as a target gene of miR-128-3p,and miR-128-3p negatively regulated the expression of SLC39A7.Over-expression of SLC39A7 partially reversed the effect of miR-128-3p on apoptosis of ovarian cancer cells with endoplasmic reticulum stress.Conclusion:miR-128-3p promotes endoplasmic reticulum stress-induced apoptosis of ovarian cancer cells by targeted down-regulation of SLC39A7.

关 键 词：卵巢癌 微RNA-128-3p SLC39A7 内质网应激 细胞凋亡 

分 类 号：R737[医药卫生—肿瘤]