摘要
目的探究shRNA-高迁移率族蛋白N2(HMGN2)慢病毒载体对肾癌细胞侵袭能力和凋亡水平的影响,并研究其作用机制。方法选择2017年1月至2018年12月天津市第四中心医院收治的31例肾细胞癌患者,术中收集患者的肿瘤组织和瘤旁组织。利用免疫组织化学染色,采用RT-qPCR和Western blot等方法检测ShRNA-HMGN2的表达以及凋亡分子标志物Caspase-3和Caspase-9的表达,利用shRNA干扰技术构建shRNA-HMGN2慢病毒载体,利用人肾癌细胞系ACHN和A498细胞作为体外实验的对象,根据有无转染shRNA-HMGN2质粒,将细胞分成三组,即A组细胞未做任何处理,B组细胞经空白载体慢病毒转染,C组细胞经shRNA-HMGN2载体构建的慢病毒转染。Tmnswell试剂盒检测三组细胞的侵袭能力和凋亡水平。结果肿瘤组织的HMGN2蛋白的表达量要明显高于瘤旁组织(P<0.05)。在人肾癌细胞系ACHN细胞和A498细胞中,C组的细胞侵袭能力要低于A组和B组(P<0.05),C组的细胞凋亡水平要高于A组和B组(P<0.05)。另外,人肾癌ACHN细胞和A498细胞中,C组HMGN2的mR-NA相对表达量和蛋白表达量要低于A组和B组(P<0.05),而C组Caspase-9和Caspase-3的mRNA相对表达量和蛋白表达量要高于A组和B组(P<0.05)。结论shRNA-HMGN2慢病毒载体可以抑制肾细胞癌的侵袭,促进肾细胞癌的凋亡,可以作为潜在的治疗靶点。
Objective To investigate the effect of shRNA-HMGN2 lentivirus vector on the invasive ability and apoptotic level of renal cell carcinoma cells and explore the mechanism.Methods Thirty-one patients with renal cell carcinoma who were admitted to the Tianjin fourth central hospital from January 2017 to December 2018 were selected for the study.Tumor tissues and paraneoplastic tissues were collected during the operation.Immunohistochemical staining,RT-qPCR and Western blot were used to detect the expression of HMGN2 as well as the expression of apoptotic molecular markers Caspase-3 and Caspase-9.The lentiviral vector of shRNA-HMGN2 was constructed by shRNA interference technology.Human renal cancer cell lines ACHN and A498 were used as experimental objects in vitro.According to whether shRNA-HMGN2 was transfected or not.The cells was divide into three groups:group A cells without any treatment,group B cells were transfected with lentivirus,group C cells were transfected with lentivirus constructed by shRNA-HMGN2 vector,Transwell kit was used to detect the invasive ability of the three groups of cells,AV-PI apoptotic reagent and the level of apoptosis.Results The expression of HMGN2 protein in tumor tissues was significantly higher than that in paraneoplastic tissues(P<0.05).In human renal cell carcinoma cell lines ACHN and A498,the invasive ability of cells in group C was significantly lower than that in group A and B(P<0.05).The apoptotic level in group C was significantly higher than that in group A and group B(P<0.05).In addition,the relative expression of HMGN2 in ACHN cells and A498 cells in group C was significantly lower than that in group A and B(P<0.05),while the relative expression of C aspase-9 and Caspase-3 in group C was significantly higher than that in group A and B(P<0.05).Conclusions The shR-NA-HMGN2 lentivirus vector can inhibit the invasion of renal cell carcinoma and promote the apoptosis of renal cell carcinoma.It can be used as a potential therapeutic target.
作者
付茂辉
孙荣凯
Fu Maohui;Sun Rongkai(Department of Urology,Tianjin Fourth Central Hospital,Tianjin 300413,China;Department of Urology,the 960th Hospital of the Joint Support Force of the Chinese People’s Liberation Army,Jinan 250031,China)
出处
《国际泌尿系统杂志》
2021年第5期794-798,共5页
International Journal of Urology and Nephrology
关键词
癌
肾细胞
高迁移率族蛋白质类
慢病毒载体
Carcinoma,Renal Cell
High Mobility Group Proteins
Lentivirus Vector
