RNA干扰组蛋白去乙酰化酶4诱导骨肉瘤细胞凋亡  

RNA interfering histone deacetylase 4 induces apoptosis of osteosarcoma cells

作　　者：肖前仁 黄路[2] 张中卒[3] 张战民[4] 陈少卿[4] 舒勇[1] 曹凯[1] 

机构地区：[1]南昌大学第一附属医院骨科,江西南昌330006 [2]江西省妇幼保健院儿童保健科,江西南昌330006 [3]重庆医科大学附属永川医院骨科,重庆402160 [4]南昌大学第一附属医院肿瘤科,江西南昌330006

出　　处：《基础医学与临床》2016年第12期1693-1698,共6页Basic and Clinical Medicine

摘　　要：目的探讨沉默组蛋白去乙酰化酶4(HDAC4)对骨肉瘤细胞HOS,U2OS生物学行为的影响。方法用脂质体瞬时转染法将化学合成HDAC4-siRNA转染至骨肉瘤HOS及U2OS细胞中,实时荧光定量PCR(q-PCR)法检测细胞中HDAC4 mRNA的表达;蛋白质免疫印迹法(Western blot)检测细胞HDAC4及其下游基因HIF-1α的蛋白表达;CCK-8法检测细胞的增殖能力;流式细胞仪检测细胞凋亡;Transwell小室法检测细胞的侵袭能力。结果转染后HOS及U2OS细胞中HDAC4 mRNA表达明显降低(P<0.01)。沉默HDAC4后HOS细胞早期凋亡率为(16.8±2.1)%,较si-control组(10.2±2.5)%明显增加(P<0.05);U2OS细胞早期凋亡率为(21.4±3.1)%,较si-control组(12.5±2.3)%明显增加(P<0.05);HOS细胞si-con组及si-HDAC4组细胞的穿膜细胞数分别为(146±34)个、(45±20)个,U2OS细胞为(207±35)个、(121±23)个,分别显著低于si-con组(P<0.05)。si-HDAC4能够明显降低HDAC4以及下游基因HIF-1α的表达。结论针对HDAC4基因的特异性RNA小干扰片段能够下调HDAC4mRNA及蛋白的表达,并抑制骨肉瘤细胞的增殖和侵袭,诱导细胞凋亡。Objective To explore the function of HDAC4 gene on biological behaviors of HOS and U2 OS osteosarcoma cells lines which inhibit HDAC4 gene expression using a small interfering RNA. Methods The HOS and U2 OS cells were transfected with a chemical synthesized small interfering RNA targeting HDAC4; the real timePCR and Western blot assays were performed to detect the mRNA and protein expression levels of HDAC4 and its downstream genes HIF-1 α,respectively,the CCK-8,flow cytometry（ FCM）,invasion assays were used to identify the cell proliferation,apoptosis and invasion abilities after the transfection with si-HDAC4,respectively.Results The q-PCR analysis showed that HDAC4 mRNA expression was down-regulated in HOS and U2 OS cells after transfected with si-HDAC4（ P〈 0. 01）; in addition,the cell proliferation was inhibited and cell apoptosis was induced by suppression of HDAC4（ P〈 0. 05）. The cells of HOS and U2 OS migrating through the membrane in si-con group and si-HDAC4 group were（ 146 ± 34）,（ 45 ± 20） and（ 207 ± 35）,（ 121 ±23）,（ P〈 0. 05）,respectively. The expression of HDAC4 and HIF-1 α was significantly reduced. Conclusions HDAC4 si-RNA can inhibit cell proliferation,invasion and induce cell apoptosis of human HOS and U2 OS osteosarcoma cells.

关 键 词：骨肉瘤 RNAI 组蛋白去乙酰化酶4 

分 类 号：R738.1[医药卫生—肿瘤]