广东医学2024，Vol.45Issue(12)：1549-1558,10.DOI:10.13820/j.cnki.gdyx.20240962
miR-200c-3p通过靶向KRAS调控ERK/AKT信号通路对肾癌细胞增殖、侵袭和迁移的影响
Effects of miR-200c-3p targeting KRAS on ERK/AKT pathway in regulating proliferation,invasion,and mi-gration of renal cancer cells
摘要
Abstract
Objective To investigate the role of miR-200c-3p in modulating the ERK/AKT signaling pathway by targeting KRAS,and its effects on the proliferation,invasion,and migration of renal cancer cells.Methods Quanti-tative real-time PCR(qRT-PCR)and Western blot were employed to assess the expression of miR-200c-3p,KRAS,and ERK/AKT-related proteins in human renal epithelial cells(HREpiC)and renal carcinoma cell lines(786-O,769-P,Caki-2).Cultured 786-O cells were divided into groups:control,miR-200c-3p mimics,si-KRAS,NC-miR-200c-3p+NC-KRAS,and miR-200c-3p mimics+pUC57-KRAS.Post-transfection,qRT-PCR and Western blot analysis were conducted to evaluate the expression levels of miR-200c-3p,KRAS,and ERK/AKT pathway proteins.Cell proliferation was assessed using EdU staining and colony formation assays;apoptosis was measured by flow cytometry,and cell migration and invasion were evaluated by Transwell assays.Additionally,mark-ers related to proliferation(Cyclin D1),apoptosis(caspase-3,cleaved caspase-3,Bax),and epithelial-mesenchy-mal transition(EMT)(E-cadherin,Vimentin)were analyzed by Western blot.A xenograft model in nude mice was used to examine tumor growth,and a dual-luciferase reporter assay was employed to verify the targeting of KRAS by miR-200c-3p.Results Compared to HREpiC cells,renal carcinoma cell lines(786-O,769-P,and Caki-2)ex-hibited significantly lower miR-200c-3p expression(P＜0.05)and higher KRAS,p-ERK1/2,and p-AKT levels(P＜0.05).In the miR-200c-3p mimics and si-KRAS groups,apoptosis rate,caspase-3,cleaved caspase-3,Bax,and E-cadherin levels were significantly higher than in the control group(P＜0.05),while KRAS expression,p-ERK1/2,p-AKT,proliferation rate,colony formation,migration,invasion,tumor volume,and Cyclin D1 and Vimentin expression were reduced(P＜0.05).In the NC-miR-200c-3p+NC-KRAS group,no significant changes were ob-served.When KRAS was reintroduced in the miR-200c-3p mimics group,apoptosis rates and associated markers de-creased while KRAS,p-ERK1/2,p-AKT,proliferation,and invasion indicators increased(P＜0.05).Conclusion miR-200c-3p downregulates KRAS expression,thereby inhibiting the ERK/AKT signaling pathway,leading to reduced proliferation,invasion,EMT,and migration of renal cancer cells while promoting apoptosis.These findings suggest miR-200c-3p as a potential target for renal cancer treatment.关键词
miR-200c-3p/Kirsten大鼠肉瘤病毒癌基因同源物/ERK/AKT/肾癌/增殖/侵袭/迁移Key words
miR-200c-3p/kirsten rats arcomaviral oncogene homolog/ERK/AKT/renal cancer/prolifera-tion/invasion/migration分类
医药卫生引用本文复制引用
陈烈钳,严秋霞,黄栋强,岳巍巍,谢进东..miR-200c-3p通过靶向KRAS调控ERK/AKT信号通路对肾癌细胞增殖、侵袭和迁移的影响[J].广东医学,2024,45(12):1549-1558,10.基金项目
广东省惠州市科技计划项目(2021WC0106267) （2021WC0106267）
