昆明医科大学学报2025，Vol.46Issue(7)：10-17,8.DOI:10.12259/j.issn.2095-610X.S20250702
环指蛋白41调控胆管癌细胞增殖和转移的生物学作用
Biological Role of RNF41 in Regulating Proliferation and Metastasis of Cholangiocarcinoma Cells
摘要
Abstract
Objective To explore the role of ring finger protein 41(RNF41)in the initiation and progression of cholangiocarcinoma.Methods The expression levels of RNF41 mRNA and protein in tumor tissues and adjacent normal tissues from 84 CHOL patients who underwent total surgical resection at the Second Affiliated Hospital of Kunming Medical University and Kunming Ganmei Hospital between January 2010 and December 2016 were analyzed using bioinformatics,Western blot,and immunohistochemistry.The TIMER 2.0 database was used to analyze the impact of RNF41 on the prognosis and survival of CHOL patients and the relationship between RNF41 and tumor clinical characteristics.RNF41 siRNA was transfected into HCC9810,RBE,and HUCCT1 cells.CCK-8,Edu,colony formation,and Western blot assays were conducted to evaluate the changes in proliferation of cholangiocarcinoma cells between the RNF41 knockdown group and the control group.Transwell assays and detection of EMT and migration markers were performed to assess changes in the invasion ability of cholangiocarcinoma cells between the RNF41 knockdown and control groups.Western blot was used to examine the effect of RNF41 knockdown on epithelial-mesenchymal transition in cholangiocarcinoma cells.Twelve BALB/c mice were randomly divided into two groups:a control group and an RNF41 knockdown group,with six mice in each group.Tumor formation assays,Western blot assays,and immunohistochemistry staining were carried out to investigate the effect of RNF41 knockdown on tumor growth in nude mice.Results Real-time quantitative fluorescence PCR analysis revealed that the expression level of RNF41 mRNA in cholangiocarcinoma tissues was significantly higher than that in the corresponding adjacent non-tumor tissues(P<0.01),and this trend was corroborated at the protein level by immunohistochemical staining.Using the TIMER 2.0 database,we further analyzed the correlation between RNF41 expression and clinicopathological features,including histological grade,tumor stage,lymph node metastasis,and patient survival.The results indicated that elevated RNF41 expression was significantly associated with advanced histological grade and lymph node metastasis in cholangiocarcinoma(P<0.01).Survival analysis demonstrated that high RNF41 expression was closely linked to poor prognosis in patients with cholangiocarcinoma(CHOL).Functional assays,including CCK-8,EdU incorporation,and colony formation,showed that RNF41 knockdown significantly inhibited the proliferation of cholangiocarcinoma cells compared with the control group.Western blot analysis revealed that,following RNF41 silencing,the expression of the epithelial-mesenchymal transition(EMT)marker E-cadherin was markedly upregulated,whereas the levels of mesenchymal markers N-cadherin and MMP9 were significantly reduced(P<0.05).These findings were consistent with the results obtained from in vitro experiments(P<0.01).Moreover,in vivo studies showed that RNF41 knockdown suppressed subcutaneous tumor formation in nude mice(P<0.05).Conclusion RNF41 plays a critical role in promoting the occurrence and progression of cholangiocarcinoma and is closely associated with adverse clinicopathological features and poor prognosis in patients.The knockdown of RNF41 effectively suppresses the proliferation,invasion,epithelial-mesenchymal transition(EMT),and tumorigenicity of cholangiocarcinoma cells.关键词
胆管癌/环指蛋白41/增殖/侵袭Key words
Cholangiocarcinoma/Ring finger protein 41/Proliferation/Invasion分类
医药卫生引用本文复制引用
吴起杰,李勇,张瑜,冉凤明,丁荣,张麒,杨银山..环指蛋白41调控胆管癌细胞增殖和转移的生物学作用[J].昆明医科大学学报,2025,46(7):10-17,8.基金项目
国家自然科学基金(82360447) （82360447）
云南省科技厅-昆明医科大学应用基础研究联合专项基金(202201AY070001-158 （）
202301AY070001-113 （）
202301AY070001-128 （）
202501AY070001-106) （）
