miR-204靶向BDNF调控上皮性卵巢癌细胞抗失巢凋亡的实验研究  

miR-204 enhances anoikis sensitivity in epithelial ovarian cancer cell line by targeting BDNF

作　　者：闫洪亮 李杨[1] 靳翠平[1] 宫晓锦 李鹏飞[1] 胡同秀 Yan Hongliang;Li Yang;Jin Cuiping(Department of Obstetrics and Gynecology,Tianjin Hospital,Tianjin 300211)

机构地区：[1]天津市天津医院妇产科

出　　处：《现代妇产科进展》2019年第9期647-650,656,共5页Progress in Obstetrics and Gynecology

基　　金：天津市天津医院科技基金(No:TJYY1801)

摘　　要：目的:探讨miR-204靶向BDNF对OV2008卵巢癌细胞抗失巢凋亡的影响及其可能机制。方法:选用OV2008细胞株,分为贴壁培养组和失巢凋亡组;qRT-PCR法检测miR-204表达水平;选择失巢凋亡组存活细胞,分为空白对照组、转染试剂组、miR-204 NC组、miR-204质粒组进行转染,qRT-PCR检测miR-204表达。应用四组细胞分别建立失巢凋亡模型,流式细胞术检测细胞凋亡率,Transwell实验检测细胞侵袭能力,qRT-PCR检测BDNF mRNA表达水平,Western blot法检测p-Akt和总Akt蛋白表达水平。结果:失巢凋亡组较贴壁培养组的miR-204含量降低( P <0.05)。与空白对照组、转染试剂组、miR-204 NC组比较,miR-204质粒组的miR-204表达水平高( P <0.05),细胞凋亡率增加( P <0.05),侵袭能力减弱( P <0.05),BDNF mRNA表达水平降低( P <0.05),p-Akt/总Akt值降低( P <0.05);前3组比较差异均无统计学意义( P >0.05)。结论: miR-204在抗失巢凋亡表型的OV2008细胞株中低表达,过表达miR-204能抑制OV2008细胞的抗失巢凋亡能力,其机制可能与下调BDNF基因表达进而抑制Akt磷酸化有关。Objective: To observe the effects of miR-204 on OV2008 cells anoikis sensitivity,and to explore possible mechanism. Methods: The OV2008 cells were divided into two groups (adhesive culture group and anoikis group),miR-204 expression level were measured respectively by qRT-PCR.Screened survival cells in anoikis group,then divided into four groups (control,mock,negative and miR-204 plasmid group),miR-204 expression level was measured by qRT-PCR.We established anoikis pattern again,apoptosis rate was measured by FACS,cell invasion ability were detected by transwell invasion assay.The mRNA level of BDNF was also detected by qRT-PCR.p-Akt expression was assessed by Western blot. Results: Expression of miR-204 was significantly down-regulated in anoikis group.Restored expression level of miR-204 enabled cells to acquire more sensitive to anoikis and decrease invasive behavior,and also resulted in BDNF down-expression and inhibited the level of Akt phosphorylation. Conclusions: Up-regulation of miR-204 maybe directly linked with the sensitivity of OV2008 cell anoikis by contributing to BDNF down-regulation,and then inhibit the level of Akt phosphorylation.

关 键 词：miR-204 上皮性卵巢癌 失巢凋亡 BDNF PI3K/AKT 

分 类 号：R737.33[医药卫生—肿瘤]