微纤丝相关蛋白4在肾透明细胞癌中的表达及其临床意义  

Expression and its clinical significance of microfibril-associated protein 4 in renal clear cell carcinoma

作　　者：杨康 程帆[1] 曾艳[1] 葛名欢[1] Yang Kang;Cheng Fan;Zeng Yan;Ge Minghuan(Department of Urology,Renmin Hospital of Wuhan University,Wuhan 430060,China)

机构地区：[1]武汉大学人民医院泌尿外科,430060

出　　处：《中华实验外科杂志》2022年第5期885-887,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金青年项目(82100703);中央高校青年教师资助项目(2042021kf0099)。

摘　　要：目的探讨微纤丝相关蛋白4(MFAP4)对肾透明细胞癌(ccRCC)的作用及其机制。方法2020年6月至2020年12月武汉大学人民医院泌尿外科收集的30对肾癌组织及细胞标本,通过实时定量免疫荧光PCR和免疫印迹方法检测MFAP4蛋白表达量;利用腺病毒构建过表达MFAP4稳转肿瘤细胞株,通过细胞迁移和侵袭实验检测MFAP4对ccRCC细胞株的影响,利用免疫荧光和免疫印迹技术检测MFAP4对ccRCC的作用机制。多组间比较采用单因素方差分析,两两比较采用t检验。结果在肾癌组织样本和肾肿瘤细胞株中MFAP4的转录水平(转录表达值:5.22±2.61比32.08±7.12,t=17.68,P<0.05)和蛋白水平表达量(条带相对灰度值:1.00±0.01比2.30±0.25,t=9.18,P<0.05)显著高于正常组。同时,MFAP4过表达组ccRCC侵袭细胞数量显著高于转染对照组(细胞数:907±63比1997±101,t=12.87,P<0.05;1208±98比2364±121,t=15.87,P<0.05),免疫荧光和免疫印迹结果提示,过表达MFAP4降低LC3B表达量(条带相对灰度值:1.00±0.01比0.65±0.08,t=46.42,P<0.05;1.00±0.01比0.67±0.06,t=26.46,P<0.05),促进p62表达(条带相对灰度值:1.00±0.01比2.27±0.04,t=7.93,P<0.05;1.00±0.01比2.17±0.08,t=9.15,P<0.05)。结论MFAP4可能通过抑制细胞自噬促进肾癌进展。Objective To investigate the effect and mechanism of microfibril-associated protein 4(MFAP4)on renal clear cell carcinoma(ccRCC)using an in vitro model.Methods Totally,30 pairs of tissue specimens of renal cancer patients collected from Department of Urology,Renmin Hospital of Wuhan University during June 2020 to December 2020,and renal cancer cell lines were used to detect the expression of MFAP4 protein by real-time quantitative immunofluorescence PCR and Western blotting.Adenovirus vector was used to construct stably transfected tumor cell lines overexpressing MFAP4,which were detected by cell migration and invasion assays.The action mechanism of MFAP4 on ccRCC was detected by immunofluorescence and Western blotting.Results The expression of MFAP4 was significantly increased in renal cancer tissue samples and renal tumor cell lines(5.22±2.61 vs.32.08±7.12,t=17.68,P<0.05;1.00±0.01 vs.2.30±0.25,t=9.18,P<0.05).Compared with the transfection control group,overexpression of MFAP4 significantly increased the number of ccRCC invasive cells(cells:907±63 vs.1997±101,t=12.87,P<0.05;1208±98 vs.2364±121,t=15.87,P<0.05),inhibited the expression of LC3B(1.00±0.01 vs.0.65±0.08,t=46.42,P<0.05;1.00±0.01 vs.0.67±0.06,t=26.46,P<0.05),and promoted the expression of p62(1.00±0.01 vs.2.27±0.04,t=7.93,P<0.05;1.00±0.01 vs.2.17±0.08,t=9.15,P<0.05).Conclusion MFAP4 may promote the progression of renal cancer by inhibiting autophagy.

关 键 词：微纤丝相关蛋白4 肾透明细胞癌 自噬 

分 类 号：R737.11[医药卫生—肿瘤]