HMGN5基因敲除对人膀胱上皮癌细胞顺铂化疗敏感性的影响  

Influence of HMGN5 gene knockout on cisplatin chemosensitivity of human bladder epithelial cancer cells

作　　者：周剑文[1] 罗广承[1] 王新君[1] 颜志坚[1] ZHOU Jian-wen;LUO Guang-cheng;WANG Xin-jun;YAN Zhi-jian(Department of Urology,Zhongshan Hospital Affiliated to Xiamen University,Xiamen 361003,China)

机构地区：[1]厦门大学附属中山医院泌尿外科

出　　处：《实用药物与临床》2019年第9期907-911,共5页Practical Pharmacy and Clinical Remedies

基　　金：厦门市科技惠民项目(3502Z20174077)

摘　　要：目的探讨HMGN5基因敲除对人膀胱上皮癌细胞顺铂化疗敏感性的影响。方法采用不同浓度CDDP(0、1、2、4,8、16μg/ml)处理人UBC细胞系,采用Western blot、菌落形成、细胞侵袭、细胞凋亡及Hoechst 33342染色法评价HMGN55蛋白敲除对UBC细胞CDDP敏感性的影响,同时分析HMGN5基因参与到UBC细胞CDDP治疗环节可能分子机制。结果 UBC细胞株5637、t24及UM-UC-3中HMGN5蛋白表达,其中5637细胞系HMGN5蛋白水平最高,而UM-UC-3细胞中最低(P<0.05);5637细胞系对CDDP敏感性显著低于其他两种(P<0.05),且UM-UC-3细胞系敏感性最高;UBC细胞系HMGN5敲除后P-Akt表达显著下降(P<0.05);CDDP处理后P-Akt活性亦随之降低(P<0.05);CDDP处理还可下调VEGF-C和SLUG表达,上调E-Cad表达(P<0.05);5637细胞HMGN5基因敲除后早期凋亡率显著提高,而加入IGF-1可逆转这一过程(P<0.05);与阴性对照组和IGF-1治疗组相比,HMGN5敲除组凋亡核百分比显著升高(P<0.05);与对照组相比,HMGN5基因敲除显著下调P-AKT和VEGF-C表达,上调E-Cad、细胞色素C、裂解半胱天冬酶-3、裂解PARP表达(P<0.05);而添加IGF-1可逆转以上蛋白质表达变化(P<0.05)。结论 HMGN5可能是顺铂治疗潜在靶点,抑制HMGN5基因有助于增加UBC细胞化疗敏感性,这一作用主要通过干扰PI3K/Akt信号通路实现。Objective To investigate the influence of HMGN5 gene knockout on cisplatin chemosensitivity of human bladder epithelial cancer cells.Methods Human UBC cell lines were treated with different concentrations of CDDP(0,1,2,4,8 and 16 μg/ml).Western blotting,colony formation,cell invasion,cell apoptosis and Hoechst 33342 staining were used to evaluate the effect of HMGN55 protein knockout on CDDP sensitivity of UBC cells.The possible molecular mechanism of HMGN5 gene involved in CDDP treatment of UBC cells was also analyzed.Results The expression of HMGN5 protein in UBC cell lines 5637,T24 and UM-UC-3 was the highest in 5637 cell lines,and was the lowest in UM-UC-3 cell lines(P<0.05).The sensitivity of 5637 cell lines to CDDP was significantly lower than that of the other two cell lines(P<0.05);UM-UC-3 cell lines had the highest level of HMGN5 protein(P<0.05).The sensitivity of P-Akt decreased significantly after HMGN5 knockout and its activity also decreased after CDDP treatment(P<0.05).CDDP treatment could also down-regulate the expression of VEGF-C and SLUG,and up-regulate the expression of E-Cad(P<0.05).The apoptosis rate of the 5637 cell after HMGN5 gene knockout significantly increased,while IGF-1 addition could reverse the process(P<0.05).The percentage of apoptotic nuclei in HMGN5 knockout group was significantly higher than that in negative control group and IGF-1 treatment group(P<0.05).HMGN5 knockout significantly decreased the expression of P-AKT and VEGF-C,and increased the expression of E-Cad,cytochrome C,lysis caspase-3 and lysis PARP(P<0.05),while the addition of IGF-1 could reverse the expression change of P-AKT and VEGF-C(P<0.05).Conclusion HMGN5 maybe the potential target for cisplatin therapy.Inhibition of HMGN5 gene can increase the chemosensitivity of UBC cells.This effect should be achieved mainly by interfering with PI3 K/Akt signaling pathway.

关 键 词：HMGN5基因 人膀胱上皮癌细胞 顺铂 化疗 敏感性 

分 类 号：R737.14[医药卫生—肿瘤]