实用医学杂志 ›› 2021, Vol. 37 ›› Issue (10): 1240-1244.doi: 10.3969/j.issn.1006⁃5725.2021.10.002

• 基础研究 • 上一篇    下一篇

视黄醇结合蛋白2基因沉默对卵巢癌SKOV3/PTX细胞迁移侵袭的影响

冯同富,陈德军, 叶常鸿, 汪黎明,金晶,雷燕, 李力   

  1. 1 湖北省妇幼保健院妇科(武汉 430070);2 广西医科大学区域性高发肿瘤早期防治研究教育部重点实验室 (南宁 530021)

  • 出版日期:2021-05-25 发布日期:2021-05-25
  • 基金资助:

    湖北省自然科学基金知识创新专项(编号 2018CFC840);湖北省卫计委联合基金(编号:WJ2018H0158);武 汉市中青年医学骨干人培养工程(编号:No.[2018]116)

Effect of RBP2 gene silencing on migration and invasion of ovarian cancer SKOV3/PTX cells 

FENG Tong⁃ fu,CHEN Dejun,YE Changhong,WANG Liming,JIN Jing,LEI Yan,LI Li.    

  1. Department of Gynecology,Mater⁃ nal and Child Health Hospital of Hubei Province,Wuhan 430070,China 

  • Online:2021-05-25 Published:2021-05-25

摘要:

目的 探讨沉默视黄醇结合蛋白 2(RBP2)基因表达对卵巢癌细胞 SKOV3/PTX 迁移及侵袭的影响及机制。方法 建立稳定干扰 RBP2 表达的细胞株 SKOV3/PTX⁃RBP2i,将实验分为 3 组,即 RBP2 沉默组(SKOV3/PTX⁃RBP2i)、阴性对照组(SKOV3/PTX⁃NC)和空白对照组(SKOV3/PTX),Western blot 法验证 RBP2基因的沉默效果,划痕实验检测三组细胞的迁移能力,Transwell 侵袭实验检测三组细胞的侵袭能力, RT⁃PCR Western blot 法检测三组细胞β⁃catenin、VEGF MMP⁃2 的表达情况。结果 成功构建稳定干扰 RBP2表达的细胞株SKOV3/PTX⁃RBP2i,沉默组RBP2的表达强度明显低于空白组和阴性对照组;沉默组、阴性对照组及空白组细胞的划痕愈合率和侵袭能力差异有统计学意义(P<0.01),沉默组细胞的划痕愈合率和侵袭能力明显低于阴性对照组和空白组(P<0.01);RT⁃PCR 和Western blot结果显示,沉默组β⁃catenin、VEGF 及MMP⁃2 的 mRNA 和蛋白表达水平均明显低于阴性对照组及空白组(P<0.05)。结论 干扰 SKOV3/PTX 细胞中RBP2的表达可抑制细胞的迁移及侵袭能力,其机制可能与Wnt/β⁃catenin 信号转导通路有关。

关键词: 卵巢肿瘤,  , 肿瘤,  , RBP2,  , 迁移,  , 侵袭

Abstract:

Objective To investigate the mechanism of RBP2 gene silencing on the migration and invasion of the ovarian cancer cell line SKOV3/PTX. Methods The cell line that stably interferes with RBP2 expression was established,and the cells were divided into three groups:RBP2 silence(SKOV3/PTX⁃RBP2i),negative control (SKOV3/PTX⁃NC),and blank control(SKOV3/PTX). Western blot was used to confirm the silencing effect of the RBP2 gene,wound healing assay was used to detect the migration ability of cells in three groups,transwell inva⁃ sion assay was used to detect the invasive ability of cells in three groups,and RT⁃PCR and western blot methods were used to detect molecular markers(β ⁃ catenin,VEGF and MMP ⁃ 2). Results The cell line SKOV3/PTX ⁃ RBP2i,which stably inhibits RBP2 expression,was successfully constructed. The RBP2 expression level of silent group was significantly lower than that of negative control group and blank group. There were significant differences in wound healing rate and invasive ability among silent group,negative control group and blank group(P<0.01). The wound healing rate and invasive ability of cells in silent group were significantly lower than those in negative control group and blank group(P<0.01). RT ⁃PCR and western blot showed that the mRNA and protein expression levels of β⁃catenin,VEGF and MMP⁃2 in the silent group had significantly lower than those in the negative control group and the blank group(P<0.05). Conclusion Interfering with RBP2 expression in SKOV3/PTX cells can prevent cell migration and invasion,and the mechanism is thought to be related to the Wnt/ β⁃catenin signaling pathway. 

Key words:

ovarian tumor, tumor, RBP2, migration, invasion