下调组蛋白去乙酰化酶表达抑制前列腺癌细胞恶性表型及其机制  

The study of suppression and action mechanism of malignant phenotype by downregulating expres. sion of histone deacetylase in prostate cancer cell

作　　者：马渊[1] 汤磊[2] 冯连成[1] 耿进成[1] 李纪夫[1] 

机构地区：[1]新疆医科大学第五附属医院泌尿外科,乌鲁木齐830011 [2]新疆生产建设兵团医院泌尿外科,乌鲁木齐830011

出　　处：《中华实验外科杂志》2017年第12期2026-2030,共5页Chinese Journal of Experimental Surgery

摘　　要：目的探讨下调组蛋白去乙酰化酶（HDAC）家族成员HDAC1和HDAC2表达抑制前列腺癌细胞恶性表型及其作用机制。方法运用脂质体2000转染试剂将siHDAC转染或HDAC1抑制剂MHY219作用LNCaP和DU145细胞，命名为LNCaP—siHDAC、DU145-siHDAC细胞和阴性对照LNCaP—NC小干扰RNA（siRNA）、DU145-NCsiRNA细胞；Westernblot检测HDAC1、HDAC2、HDAC3、基质金属蛋白酶（MMP）-1、MMP-2、MMP-9、基质金属蛋白酶抑制因子（TIMP）-1蛋白表达，RealtimeRT—PCR检测MMP-1、MMP-2、MMP-9、TIMP-1mRNA的表达水平。噻唑蓝（M．rr）、克隆形成实验及Transwell法检测细胞活性、克隆形成能力和迁移率。结果LNCaP—siHDAC和DU145．siHDAC细胞中HDAC1和HDAC2相对表达量分别为0．22±0．Ol、0．25±0．02和0．15±0．叭、0．30±0．03，显著低于NCsiRNA组1．00±0．05和1．00±0．03（F㈨。P=289．440，P=0．008，FDU145=336．125，P=0．002）；MHY219处理的LNCaP和DU145细胞中HDAC1和HDAC2相对表达量分别为0．53±0．04、0．60±0．02和0．58±0．04、0．66±0．05，显著低于Untreated组（1．00±0．03）（FLNCaP=147．158，P=0．022；FDJ145=134．573，P=0．035）。LNCaP—siHDAC和DU145-siHDAC细胞活性分别为（40．38±4．16）％和（21．42±5．26）％，与NCsiRNA组（98．47±6．70）％和（99．15±7．00）％比较差异有统计学意义（FlncAp=125．627，P=0．004；FDu145=137．560，P=0．000）；MHY219处理的LNCaP和DU145细胞活性分别为（61．75±7．12）％和（45．00±5．53）％，显著低于Untreated组（99．34±7．12）％和（98．58±8．05）％（FLNCAph=88．392，P=0．032；FDU145=103．10，P=0．017）。LNCaP—siHDAC和DU145-siHDAC细胞克隆形成率分别为（26．37±3．50）％和（19．48±2．33）％，与NCsiRNA组（97．20±7．88）％和（99．23±9．52）％比较差异有统计学意义（F㈨。P=287，653，P=0，000；FnU。=315．042，P=0．000）；MHY219处理的LNCaP和DU145细胞Objective Histone deacetylase （HDAC） promoted growth and metastasis of prostate cancer cell and it＇ s action mechanism, however, remain unclear. The aim of present study is investigate the suppression and action mechanism of malignant phenotype by downregulating expression of histone deacetylase （ HDAC1 and HDAC2） in prostate cancer cell. Methods Prostate cancer cell lines, LNCaP and DU145 cells were transfected with siHDAC using liposome 2000 or treated with MHY219, named as LNCaP- siHDAC, DU145 -siHDAC cells negative control LNCaP- NC small interfering RNA （siRNA）, DU145- NCsiRNA cells, and the expressions of HDAC1, HDAC2, HDAC3, matrix metalloproteinase （MMP） - 1, MMP - 2, MMP - 9 and TIMP - 1 protein were determined by Western blotting and the levels of MMP - 1, MMP - 2, MMP - 9 and TIMP - 1 mRNA were detected using Real time RT - PCR. Cell via- bility, clone formation ability and metastasis were monitored using MTF, clone formation assay and Tran- swell respectively in LNCaP and DU145 cells. Results The relative expression levels of HDAC1 and HDAC2 protein was respectively 0. 22 ± 0.01, 0. 25 ± 0. 02 in LNCaP - siHDAC cell and 0. 15 ± 0. 01, 0. 30 ± 0. 03 in DU145 - siHDAC celI, and significantly lower than LNCaP - NCsiRNA （ 1.00 ± 0. 05, FLNCae = 289. 440, P = 0. 008） and DU145 - - NCsiRNA （ 1.00 ± 0.03, FDU145 = 336. 125, P = 0. 002）. The relative expression levels of HDAC1 and HDAC2 protein was respectively 0. 53 ± 0. 04, 0. 60 ± 0.02 in LNCaP cell and 0. 58 ± 0. 04, 0. 66 ± 0.05 in DU145 cell treated with MHY219, and significantly lower than Untreated group （FLNCaP = 147. 158, P =0. 022 ;FDuI45 = 134. 573, P = 0. 035）. The results from the MTI＂ indicated that cell viability was respectively （40. 38 ± 4. 16 ）% in LNCaP- siHDAC cell and （21.42 ± 5.26） % in DU145 - siHDAC cell with statistical differences compared with LNCaP - NCsiRNA cell [（98.47 ± 6.70）%, FLNCaP = 125.627, P= 0.004] and DU145 - NCsiRNA cell [（99. 15 ± 7. 00）%, FDu145 = 137. 560

关 键 词：前列腺癌 组蛋白去乙酰化酶 基质金属蛋白酶 恶性表型 作用机制 

分 类 号：R737.25[医药卫生—肿瘤]