摘要
目的研究RNA结合基序单链相互作用蛋白3(RBMS3)对宫颈癌细胞增殖和转移的影响,并探讨其发挥作用的可能机制。方法采用GEPIA网站分析RBMS3在宫颈癌组织中的表达情况。qRT-PCR和Western blotting检测RBMS3 mRNA和蛋白在宫颈癌组织和细胞系中的表达水平。构建RBMS3过表达慢病毒,感染宫颈癌细胞系Caski,分为NC组和oe-RBMS3组,采用MTS检测各组细胞增殖能力,Transwell实验检测各组细胞转移能力,移植瘤实验检测各组细胞体内成瘤能力。Western blotting检测各组细胞中Wnt/β-catenin信号通路关键蛋白wnt3a、β-catenin及其下游靶蛋白cMYC、cyclinD1、MMP-2的表达。结果GEPIA网站分析结果显示RBMS3 mRNA在宫颈癌组织中的表达显著低于在正常宫颈组织中的表达。qRT-PCR和Western blotting结果均显示RBMS3 mRNA和蛋白在宫颈癌组织中的表达显著低于其在癌旁组织中的表达,RBMS3 mRNA和蛋白在宫颈癌细胞系中的表达显著低于其在人鳞状上皮永生化细胞H8中的表达。与NC组相比,oe-RBMS3组Caski细胞增殖、转移和体内成瘤能力均降低,细胞中wnt3a、β-catenin、cMYC、cyclinD1、MMP-2蛋白表达均降低。结论RBMS3可能通过调控Wnt/β-catenin信号通路抑制宫颈癌细胞增殖和转移能力。
Objective To study the effect of RNA binding motif single stranded interacting protein 3(RBMS3)on the proliferation and metastasis of cervical cancer cells,and to explore its potential mechanism.Methods GEPIA website was used to analyze the expression of RBMS3 in cervical cancer tissues.qRT-PCR and western blotting were conducted to detect the expression levels of RBMS3 mRNA and protein in cervical cancer tissues and cell lines.We constructed RBMS3 overexpression lentivirus and infected cervical cancer cell line Caski.The cells were divided into the NC group and oe-RBMS3 group.MTS was performed to detect cell proliferation ability of each group.Transwell assay was used to detect cell metastasis ability of each group,and transplanted tumor experiment was applied to detect cell tumor-forming ability in vivo.Western blotting was conducted to detect the expression levels of of key proteins wnt3a,β-catenin and their downstream target proteins cMYC,cyclinD1,and MMP-2 in the Wnt/β-catenin signaling pathway in each group of cells.Results The results of the GEPIA website analysis showed that the expression of RBMS3 mRNA in cervical cancer tissues was significantly lower than that in normal cervical tissues.Both qRT-PCR and western blotting results showed that the expression of RBMS3 mRNA and protein in cervical cancer tissues were significantly lower than those in paracancerous tissues,and the expression of RBMS3 mRNA and protein in cervical cancer cell lines was significantly lower than that in human squamous epithelial immortalized cells H8.Compared with the NC group,Caski cell’s proliferation,metastasis and tumor formation ability in the oe-RBMS3 group were reduced,and the expressions of wnt3a,β-catenin,cMYC,cyclinD1,and MMP-2 proteins in the cells were all reduced.Conclusion RBMS3 may inhibit the proliferation and metastasis of cervical cancer cells by regulating the Wnt/β-catenin signaling pathway.
作者
霍志平
李红霞
王宏利
孟宪宁
HUO Zhiping;LI Hongxia;WANG Hongli;MENG Xianning(Department of Gynecology,Hebei Petrochina Central Hospital,Langfang 065000,China)
出处
《标记免疫分析与临床》
CAS
2021年第7期1228-1234,共7页
Labeled Immunoassays and Clinical Medicine
基金
河北省2019年医学科学研究重点课题(编号:Y20190073)
廊坊市科技计划项目(编号:2017013145)。
