激活的蛋白激酶C受体1在膀胱癌组织中表达及对膀胱癌阿霉素耐药的影响及其机制  

Expression of receptor for activated c-kinase 1 in bladder cancer and its effect on adriamycin resistance in bladder cancer and its mechanism

作　　者：杨阳[1] 张刘辉[1] 樊鑫 杨小明[1] 裴圆芳 魏晓松 Yang Yang;Zhang Lihui;Fan Xin;Yang Xiaoming;Pei Yuanfang;Wei Xiaosong(Department of Urology,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,China;Department of Ear,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,China)

机构地区：[1]郑州大学第一附属医院泌尿外科,郑州450000 [2]郑州大学第一附属医院耳科,郑州450000

出　　处：《中华实验外科杂志》2023年第6期1023-1026,共4页Chinese Journal of Experimental Surgery

摘　　要：目的:探讨激活的蛋白激酶C受体1(RACK1)在膀胱癌组织中表达及对膀胱癌阿霉素耐药的影响及机制。方法:选取郑州大学第一附属医院2020年1月至2022年1月收治的37例阿霉素耐药和41例非耐药的膀胱癌组织作为研究对象,采用免疫组织化学分析耐药和非耐药膀胱癌组织RACK1蛋白表达水平。人膀胱移行细胞癌细胞系HT-1197采用梯度递增法建立阿霉素耐药细胞株HT-1197/ADM。采用蛋白质印迹法(Western blot)分析HT-1197/ADM和HT-1197细胞株RACK1蛋白表达水平。采用对照慢病毒和RACK1短发卡RNA(shRNA)慢病毒感染析HT-1197/ADM,建立析HT-1197/ADM组和析HT-1197/ADM/RACK1 KD组,采用细胞计数试剂盒(CCK-8)、EdU染色和体外移植瘤分析两组细胞增殖的能力。采用流式细胞仪分析两组细胞凋亡的影响;Western blot分析两组细胞凋亡相关蛋白表达水平。计量资料比较采用t检验。结果:耐药膀胱癌组织RACK1蛋白表达水平(9.29±1.40)明显高于非耐药膀胱癌组织(6.20±0.95),差异有统计学意义(t=11.520,P<0.05)。耐药膀胱癌细胞RACK1蛋白表达水平(1.71±0.19)明显高于正常膀胱癌细胞(1.22±0.11),差异有统计学意义(t=5.483,P<0.05)。HT-1197/ADM组细胞吸光度(A)值(1.78±0.13)明显高于HT-1197/ADM/RACK1 KD组(1.31±0.03),差异有统计学意义(t=8.340,P<0.05)。HT-1197/ADM组细胞A值(74.87±6.00)明显高于HT-1197/ADM/RACK1 KD组(54.18±7.88),差异有统计学意义(t=5.115,P<0.05)。HT-1197/ADM组细胞成瘤体积[(608.33±65.13)mm 3]明显高于HT-1197/ADM/RACK1 KD组[(487.67±54.09)mm 3],差异有统计学意义(t=3.491,P<0.05)。HT-1197/ADM组细胞成瘤质量[(4.88±0.64)g]明显高于HT-1197/ADM/RACK1 KD组[(3.46±0.57)g],差异有统计学意义(t=4.045,P<0.05)。HT-1197/ADM组细胞凋亡比例[(15.53±1.78)%]明显低于HT-1197/ADM/RACK1 KD组[(25.94±3.33)g],差异有统计学意义(t=6.746,P<0.05)。HT-1197/ADM组细胞半胱氨酰天冬氨酸特异性蛋白酶(Caspase)-3和B细胞淋�Objective To investigate the expression of receptor for activated c-kinase 1(RACK1)in bladder cancer and its effect on adriamycin resistance in bladder cancer.Methods A total of 37 cases of doxorubicin resistant and 41 cases of no drug-resistant bladder cancer tissues treated in our hospital from January 2020 to January 2022 were selected as research objects.The expression level of RACK1 protein in drug resistant and no drug resistant bladder cancer tissues was analyzed by immunohistochemistry.A human bladder transitional cell carcinoma cell line HT-1197 was established using a gradient method to develop an adriamycin resistant cell line HT-1197/ADM.The expression level of RACK1 protein in HT-1197/ADM and HT-1197 cell lines were analyzed by Western blotting.Using control lentivirus and RACK1 short hairpin RNA(shRNA)lentivirus infection to analyze HT-1197/ADM,the HT-1197/ADM group and HT-1197/ADM/RACK1 KD group were established.The cell proliferation ability of the two groups was analyzed using cell counting kit-8(CCK-8)assay,EdU staining,and in vitro tumor transplantation.The effects of apoptosis in both groups were analyzed by flow cytometry.The expression level of apoptosis related proteins in both groups was analyzed by Western blotting.Measurement data were compared using a t-test.Results The expression level of RACK1 protein in drug resistant bladder cancer tissue(9.29±1.40)was significantly higher than that in no drug resistant bladder cancer tissue(6.20±0.95,t=11.520,P<0.05).The expression level of RACK1 protein in drug-resistant bladder cancer cells(1.71±0.19)was significantly higher than that in normal bladder cancer cells(1.22±0.11,t=5.483,P<0.05).The absorbance(A)value(1.78±0.13)of cells in the HT-1197/ADM group was significantly higher than that in the HT-1197/ADM/RACK1 KD group(1.31±0.03,t=8.340,P<0.05).The A value of cells in the HT-1197/ADM group(74.87±6.00)was significantly higher than that in the HT-1197/ADM/RACK1 KD group(54.18±7.88,t=5.115,P<0.05).The tumor formation volume of HT-1197/A

关 键 词：蛋白激酶C受体1 膀胱癌 阿霉素 耐药 凋亡 

分 类 号：R737.14[医药卫生—肿瘤]