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
丁勇杰 1朱卫 2刘涛 3谢冲 2李艳松 1胡祎舜 1刘敏豪 1汤晨昊 1刘子阳 1彭袈豪 1袁作洈 1柳栩然 1程乔宇 1廖义翔 1曾金敏1
作者信息
- 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
- 折叠
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.
Abstract
Objective This study aims to investigate the effect of PSMD7 expression on the prolif-eration 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 exper-imental 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 clo-ning 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 carcino-ma.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 de-creased,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 Tran-swell 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 ability of clear cell renal cell carcinoma.
关键词
蛋白酶体26S亚基非ATP酶7/肾透明细胞癌/短发夹RNAKey words
Proteasome 26S subunit,non-ATPase 7/Clear cell renal cell carcinoma/Short hairpin RNA引用本文复制引用
出版年
2024
NETL
NSTL
万方数据