过表达内质网蛋白29抑制人卵巢癌细胞系OVCAR3侵袭和迁移

基础医学与临床 ›› 2021, Vol. 41 ›› Issue (10): 1434-1439.

过表达内质网蛋白29抑制人卵巢癌细胞系OVCAR3侵袭和迁移

董莉丽^1*, 冯磊¹, 杨然¹, 李松¹, 饶玉梅²

1.南阳市中心医院妇科,河南南阳 473000;
2.郑州大学第一附属医院妇科,河南郑州 450000

收稿日期:2020-09-15 修回日期:2021-03-13 发布日期:2021-09-29
通讯作者: *43238389@qq.com
基金资助:
河南省高等学校重点科研项目(18A310031)

Over-expression of ERP29 inhibits invasion and migration of human ovarian cancer cell line OVCAR3

DONG Li-li^1*, FENG Lei¹, YANG Ran¹, LI Song¹, RAO Yu-mei²

1. Department of Gynecology, Nanyang Central Hospital, Nanyang 473000;
2. Department of Gynecology, the First Affiliated Hospital, Zhengzhou University, Zhengzhou 450000, China

Received:2020-09-15 Revised:2021-03-13 Published:2021-09-29
Contact: *43238389@qq.com

摘要/Abstract

摘要： 目的探究内质网蛋白29(ERP29)过表达对人卵巢癌细胞系OVCAR3的侵袭、迁移能力的影响及机制。方法取对数增殖期OVCAR3细胞系,分为过表达组、空载组及对照组。过表达组用脂质体转染法转染ERP29过表达RNA重组真核载体pcDNA3.1-ERP29质粒,空载组转染阴性对照载体pcDNA3.1质粒,对照组不做处理。倒置荧光显微镜观察转染效率;用RT-qPCR检测ERP29 mRNA表达量;用Transwell小室法检测侵袭;用划痕实验检测迁移;用Western blot检测蛋白激酶B(AKT)、p-AKT、雷帕霉素靶蛋白(mTOR)、p-mTOR、真核生物始动因子4E结合蛋白1(4EBP1)、p-4EBP1蛋白表达量。结果过表达组和空载体组质粒转染效率均超过80%;与对照组和空载组相比,过表达组ERP29 mRNA相对表达量升高(P＜0.05),穿膜细胞数减少(P＜0.05),迁移率降低(P＜0.05),p-AKT/AKT、p-mTOR/mTOR、p-4EBP1/4EBP1值均降低(P＜0.05)。结论 ERP29过表达可显著抑制卵巢癌细胞系OVCAR3的侵袭、迁移能力。这可能是通过下调AKT、mTOR、4EBP1磷酸化发挥作用。

关键词: 卵巢癌, 内质网蛋白29, 侵袭, 迁移

Abstract: Objective To explore the effect of over-expression of endoplasmic reticulum protein 29 (ERP29) on the invasion and migration of human ovarian cancer OVCAR3 cells, and potentially related mechanisms. Methods The OVCAR3 cell line at logarithmic proliferation stage was divided into over-expression group, empty vector group and control group. The over-expression group was transfected with ERP29 over-expression RNA recombinant eukaryotic vector pcDNA3.1-ERP29 plasmid by liposome transfection method. The empty vector group was transfected with the negative control vector pcDNA3.1 empty plasmid without any treatment. The transfection efficiency was examined by inverted fluorescence microscopy, The expression of ERP29 mRNA after transfection was measured by RT-qPCR. Transwell chamber experiment was used to detect the invasion. The scratch experiment was used to detect the migration. Western blot was used to detect the expression of AKT, p-AKT, mTOR, p-mTOR, 4EBP1, p-4EBP1 protein. Results The plasmid transfection efficiency in over- expression group and empty vector group all exceeded 80%. Compared with the control group and the empty vector group, the relative expression of ERP29 mRNA in the over-expression group was increased (P＜0.05), the number of transmembrane cells ,migration rate and p-AKT/AKT, p-mTOR/mTOR, p-4EBP1/4EBP1 were all decreased(P＜0.05). Conclusions Over-expression of ERP29 can significantly inhibit the invasion and migration of ovarian cancer OVCAR3 cells,the underline mechanism is potentially the inhibition by down-regulating phosphorylation of AKT, mTOR, and 4EBP1.

Key words: ovarian cancer, endoplasmic reticulum protein 29, invasion, migration

中图分类号:

R737.31

董莉丽, 冯磊, 杨然, 李松, 饶玉梅. 过表达内质网蛋白29抑制人卵巢癌细胞系OVCAR3侵袭和迁移[J]. 基础医学与临床, 2021, 41(10): 1434-1439.

DONG Li-li, FENG Lei, YANG Ran, LI Song, RAO Yu-mei. Over-expression of ERP29 inhibits invasion and migration of human ovarian cancer cell line OVCAR3[J]. Basic & Clinical Medicine, 2021, 41(10): 1434-1439.

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