肿瘤抑制基因p53通过热休克蛋白B7抑制前列腺癌增殖、迁移从而抑制前列腺癌的进展  

Regulation of heat shock protein family B(small)member 7 by p53 inhibits prostate cancer proliferation,migration and progression

作　　者：张二伟[1] 李冠儒[1] 李炎生[1] 王启[1] Zhang Erwei;Li Guanru;Li Yansheng;Wang Qi(Department of Urology,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,450000

出　　处：《中华实验外科杂志》2022年第7期1345-1348,共4页Chinese Journal of Experimental Surgery

摘　　要：目的探讨热休克蛋白B7(HSPB7)对前列腺癌增殖、迁移的影响及其作用机制。方法使用肿瘤基因组图谱(TCGA)和基因表达数据库(GEO)数据库比较2006年到2021年525例前列腺癌和83例正常组织中基因HSPB7的差异表达。将人前列腺癌细胞系22RV1和DU145分为对照组和实验组,分别转染空质粒和HSPB7质粒,转染2 d后,采用细胞计数试剂盒(CCK-8)法、平板克隆形成实验、5-乙炔基-2’脱氧尿嘧啶核苷(EdU)染色实验验证细胞的增殖能力;采用划痕愈合实验和Transwell实验验证细胞的迁移能力,采用蛋白质印迹法分析去基化药物5-Aza-dC以及抑癌基因p53与HSPB7的靶向关系。两实验组间比较采用独立样本t检验,多实验组间比较采用方差分析。结果对照组细胞EdU着色细胞比例高于实验组[22RV1:(13.250±0.508)%比(7.439±0.172)%,t=10.840,P<0.05;DU145:(31.280±0.680)%比(7.708±0.369)%,t=30.460,P<0.05]、培养96 h后吸光度高于实验组(22RV1:1.742±0.563比1.315±0.651,F=16.520,P<0.05;DU145:1.960±0.443比1.578±0.469,F=28.850,P<0.05)、克隆形成数高于实验组(22RV1:407.700±5.548比164.700±7.839,t=25.300,P<0.05;DU145:503.700±6.438比301.300±5.783,t=23.380,P<0.05)、划痕愈合率高于实验组[22RV1:(20.690±0.8903)%比(3.458±0.6731)%,t=15.440,P<0.05;DU145:(51.620±2.460)%比(29.400±2.508)%,t=6.323,P<0.05]、迁移细胞数高于实验组(22RV1:78.250±7.420比23.750±2.869,t=6.851,P<0.05;DU145:218.000±6.285比58.000±5.492,t=19.170,P<0.05)。抑癌基因P53过表达组HSPB7蛋白表达量高于对照组(22RV1:1.025±0.020比1.999±0.057,t=16.130,P<0.05;DU145:1.043±0.040比2.350±0.057,t=18.870,P<0.05)。结论HSPB7在前列腺癌组织中被甲基化从而导致表达下调,HSPB7通过抑制前列腺癌细胞增殖和迁移从而抑制前列腺癌的进展,且这种作用受到抑癌基因p53的调控。Objective To investigate the effect and molecular mechanism of heat shock protein family B(small)member 7(HSPB7)on the proliferation and migration of prostate cancer.Methods Differential expression of HSPB7 in 525 cases of prostate cancer tissues and 83 cases of normal prostate tissues was compared using TCGA and GEO databases from 2006 to 2021.The human prostate cancer cell lines 22RV1 and DU145 were divided into control and experimental groups and transfected with empty plasmid or HSPB7 plasmid,respectively.After 2 days,the proliferation ability of the cells was verified using the cell counting kit-8(CCK-8)method,plate colony formation assay,and 5-Ethynyl-2′-deoxyuridine(EdU)assay.The migration ability of the cells was verified using a scratch healing assay and Transwell assay.Western blotting was used to analyze the relationship between the deglycosylated drug 5-Aza-dC and the tumor suppressor gene p53 or HSPB7.Independent sample t-test was used for comparison between two experimental groups,and analysis of variance was used for comparison between multiple experimental groups.Results The proportion of EdU-stained cells[22RV1:(13.250±0.508)%,(7.439±0.172)%,t=10.840,P<0.05;DU145:(31.280±0.680)%,(7.708±0.369)%,t=30.460,P<0.05],absorbance(22RV1:1.742±0.563,1.315±0.651,F=16.520,P<0.05;DU145:1.960±0.443,1.578±0.469,F=28.850,P<0.05),number of colony formation 22RV1:407.7±5.548,164.7±7.839,t=25.300,P<0.05;DU145:503.7±6.438,301.3±5.783,t=23.380,P<0.05),scratch healing rate[22RV1:(20.690±0.8903)%,(3.458±0.6731)%,t=15.440,P<0.05;DU145:(51.620±2.460)%,(29.400±2.508)%,t=6.323,P<0.05],and number of migrated cells(22RV1:78.25±7.42,23.75±2.869,t=6.851,P<0.05;DU145:218±6.285,58±5.492,t=19.170,P<0.05)were higher in the control group than in the experimental group.HSPB7 protein expression was higher in the tumor suppressor gene P53 overexpression group than in control group(22RV1:1.025±0.020 vs.1.999±0.057,t=16.130,P<0.05;DU145:1.043±0.040 vs.2.350±0.057,t=18.870,P<0.05).Conclusion HSPB7 is methylated i

关 键 词：前列腺癌 热休克蛋白B7 P53 甲基化 

分 类 号：R737.25[医药卫生—肿瘤]