摘要
目的 探讨微小RNA-10a(miR-10a)通过靶向调控血小板反应蛋白2/基质金属蛋白酶(THBS2/MMPs)信号通路在宫颈癌中的作用机制。方法 收集了2020年2月—2022年10月在唐山市中心医院治疗的68例宫颈癌患者样本,采用免疫组化和Western blot方法检测癌组织和癌旁正常组织中THBS2蛋白表达情况。利用双荧光素酶报告实验验证miR-10a与THBS2的靶向调控。将miR-10a、THBS2及阴性对照质粒转染至Ca Ski细胞,细胞分为阴性对照组、THBS2过表达组、miR-10a过表达组及共同过表达组。通过qRT-PCR和Western blot方法分析miR-10a对THBS2的调控作用,并应用CCK-8和Transwell实验评估miR-10a及THBS2对Ca Ski细胞增殖、迁移和侵袭的影响。qRT-PCR和Western blot方法检测细胞外基质蛋白(ECM)成分及基质金属蛋白酶2(MMP2)、MMP9的调控。利用小鼠移植瘤模型观察miR-10a对肿瘤生长的影响。结果 与癌旁正常组织比较,宫颈癌组织中THBS2表达下降(P<0.05)。miR-10a可靶向结合THBS2的3′-UTR。与阴性对照组比较,过表达mi R-10a可降低CaSki细胞中THBS2的表达水平并促进细胞增殖、迁移和侵袭(P<0.05),过表达THBS2则抑制这些过程(P<0.05)。共同过表达miR-10a和THBS2可部分抵消miR-10a的促癌效应。此外,miR-10a还下调ECM的表达(P<0.05),并上调MMP2和MMP9的表达水平(P<0.05),而THBS2则产生相反效应(P<0.05)。在小鼠移植瘤模型中,过表达miR-10a可抑制THBS2的表达水平并促进肿瘤生长(P<0.05)。结论 miR-10a通过靶向抑制THBS2及其下游THBS2/MMPs信号通路,促进宫颈癌细胞的增殖、迁移、侵袭和肿瘤瘤生长。
Objective To investigate the mechanism by which miR-10a promotes cervical cancer development by targeting the THBS2/MMPs signaling pathway.Methods Cervical cancer samples from 68 patients treated at Tangshan Central Hospital between February 2020 and October 2022 were collected.THBS2 protein expression in both cancerous and adjacent normal tissues was assessed using immunohistochemistry and Western blot analysis.The targeting relationship between miR-10a and THBS2 was confirmed via a dual-luciferase reporter assay.CaSki cells were transfected with miR-10a,THBS2,or negative control plasmids,forming four groups:negative control,THBS2 overexpression,miR-10a overexpression,and co-overexpression.The regulatory effect of miR-10a on THBS2 expression was analyzed through qRT-PCR and Western blot.CCK-8 and Transwell assays were used to evaluate the impact of miR-10a and THBS2 on CaSki cell proliferation,migration,and invasion.Additionally,qRT-PCR and Western blot were used to analyzed extracellular matrix(ECM)components and the expression of matrix metalloproteinases MMP2 and MMP9 were investigated.A mouse xenograft model was employed to assess the effect of miR-10a on tumor growth.Results THBS2 expression was significantly reduced in cervical cancer tissues compared to adjacent normal tissues(P<0.05).miR-10a was found to target the 3′-UTR of THBS2.miR-10a overexpression significantly decreased THBS2 mRNA and protein levels,promoting CaSki cell proliferation,migration,and invasion(P<0.05).In contrast,THBS2 overexpression inhibited these processes(P<0.05).Co-overexpression of miR-10a and THBS2 partially mitigated the oncogenic effects of miR-10a.Furthermore,miR-10a downregulated ECM components(P<0.05)and upregulated MMP2 and MMP9 expression(P<0.05),while THBS2 exhibited the opposite effects(P<0.05).In the mouse xenograft model,miR-10a overexpression suppressed THBS2 expression and promoted tumor growth(P<0.05).Conclusion miR-10a promotes the proliferation,migration,invasion,and tumor growth of cervical cancer cells by directly targeting and inhibiting THBS2 and modulating the downstream THBS2/MMPs signaling pathway.These findings provide new insights into the molecular mechanisms of cervical cancer,suggesting that MiR-10a may serve as a potential therapeutic target.
作者
王阳阳
王国经
刘耘
程义杰
刘艺文
WANG Yangyang;WANG Guojing;LIU Yun;CHENG Yijie;LIU Yiwen(Department of Gynaecology,Tangshan Central Hospital,Tangshan,067000,China)
出处
《中国煤炭工业医学杂志》
2024年第5期469-475,共7页
Chinese Journal of Coal Industry Medicine
基金
河北省2021年度医学科学研究课题(编号:20210127)
