蛋白酶体26S亚基非ATP酶7对肾透明细胞癌增殖和迁移能力的影响  

Effect of proteasome 26S subunit,non-ATPase 7 on bladder cancer proliferation and migration

作　　者：丁勇杰 朱卫[2] 刘涛[3] 谢冲 李艳松 胡祎舜 刘敏豪 汤晨昊 刘子阳 彭袈豪 袁作洈 柳栩然 程乔宇 廖义翔[1] 曾金敏[1] Ding Yongjie;Zhu Wei;Liu Tao;Xie Chong;Li Yansong;Hu Yishun;Liu Minhao;Tang Chenhao;Liu Ziyang;Peng Jiahao;Yuan Zuowei;Liu Xuran;Cheng Qiaoyu;Liao Yixiang;Zeng Jinmin(Department of Urology,Jingzhou Central Hospital,Jingzhou Hospital Affliated to Yangize Unwersity,Jingzhou 434020,China;Department of Urology,Jiangling County People’s Hospital,Jiangling 434100,China;Department of Urology,Wuhan University Zhongnan Hospital,Wuhan 430071,China)

机构地区：[1]荆州市中心医院长江大学附属荆州医院泌尿外科,荆州434020 [2]江陵县人民医院泌尿外科,江陵434100 [3]武汉大学中南医院泌尿外科,武汉430071

出　　处：《中华实验外科杂志》2024年第4期788-790,共3页Chinese Journal of Experimental Surgery

摘　　要：目的本研究旨在探讨蛋白酶体26S亚基非ATP酶7(PSMD7)表达对肾透明细胞癌增殖和迁移能力的影响。方法利用临床生信之家数据库分析PSMD7在肾透明细胞癌中的差异表达。在人肾透明细胞癌皮肤转移细胞(Caki-1)细胞株中利用成簇的规律间隔短回文重复序列(CRISPR)/Cas9技术导入短发夹RNA(shRNA)敲低PSMD7,设置组别为Caki-1对照组细胞株,稳定敲低PSMD7实验组的Caki-1细胞株(shPSMD7-1、shPSMD7-2),以蛋白免疫印迹法(Western blot)检测Caki-1细胞中PSMD7的敲低效率。通过细胞计数盒(CCK-8)增殖实验、平板克隆实验和Transwell实验评估实验组和对照组细胞增殖和迁移的能力。两组间比较采用独立样本t检验。结果PSMD7在多种肿瘤中表达显著上调,其中包括肾透明细胞癌,肿瘤组织中PSMD7表达高于正常组织[5.70(4.13~7.29)比3.46(1.74~5.17),P<0.05]。通过蛋白质免疫检测法检测敲低效率(1.36±0.05比0.43±0.07,t=10.77,P<0.05;1.36±0.05比0.31±0.07,t=12.53,P<0.05);CCK-8结果显示,敲低PSMD7后,细胞增殖能力显著下降(1.32±0.91比0.67±0.49,t=3.448,P<0.05;1.32±0.91比0.60±0.35,t=3.222,P<0.05)。平板克隆形成实验显示,PSMD7敲低组细胞的增殖水平明显低于对照组[(792.0±24.0)个比(520.0±28.0)个,t=7.376,P<0.05;(792.0±24.0)个比(518.0±30.0)个,t=7.132,P<0.05]。Transwell实验显示,PSMD7敲低组细胞的迁移能力明显低于对照组[(690.0±14.0)个比(237.5±33.5)个,t=12.46,P<0.05;(690.0±14.0)个比(176.5±13.5)个,t=26.40,P<0.05]。结论在肾癌组织中,PSMD7表达上调;敲低PSMD7表达可有效抑制肾癌细胞的增殖和迁移能力。Objective This study aims to investigate the effect of PSMD7 expression on the proliferation and migration ability of clear cell renal cell carcinoma.Methods The differential expression and prognosis of PSMD7 in clear cell renal cell carcinoma were analyzed using Gepia and Timer databases.Short hairpin RNA(shRNA)was introduced into the Caki-1 cell line to knock down PSMD7 using CRISPR/Cas9 technology,then the experimental grouping was set as Caki-1 control cell line and the experimental group of stabilized knockdown of PSMD7(shPSMD7-1,shPSMD7-2).The knockdown efficiency of PSMD7 in Caki-1 cells was determined by Western blotting.The proliferation and migration abilities of the experimental and control groups were assessed by cell counting kit(CCK-8)proliferation assay,plate cloning assay,and Transwell assay.Comparisons between the two groups were made using an independent samples t-test.Results PSMD7 expression was significantly upregulated in clear cell renal cell carcinoma.The expression of PSMD7 was higher in tumor tissues than in normal tissues[5.7(4.13-7.29)vs.3.46(1.74-5.17),P<0.05].The efficiency of PSMD7 knockdown was determined by Western blotting(1.36±0.05 vs.0.43±0.07,t=10.77,P<0.05;1.36±0.05 vs.0.31±0.07,t=12.53,P<0.05).Following PSMD7 knockdown,the proliferative capacity of PSMD7 knockdown cells significantly decreased,as shown by the CCK-8 results(1.32±0.91 vs.0.67±0.49,t=3.448,P<0.05;1.32±0.91 vs.0.60±0.35,t=3.222,P<0.05).Additionally,the colony formation ability of PSMD7 knockdown cells was significantly reduced,as demonstrated by the plate colony formation assay(792.0±24.0 vs.520.0±28.0,t=7.376,P<0.05;792.0±24.0 vs.518.0±30.0,t=7.132,P<0.05).The Transwell assay revealed a significant decrease in cell migration ability following PSMD7 knockdown(690.0±14.0 vs.237.5±33.5,t=12.46,P<0.05;690.0±14.0 vs.176.5±13.5,t=26.4,P<0.05).Conclusion The study found that PSMD7 was upregulated in clear cell renal cell carcinoma.Knockdown of PSMD7 effectively inhibited the proliferation and migration

关 键 词：蛋白酶体26S亚基非ATP酶7 肾透明细胞癌 短发夹RNA 

分 类 号：R737.11[医药卫生—肿瘤]