基础医学与临床 ›› 2016, Vol. 36 ›› Issue (12): 1693-1698.

• 研究论文 • 上一篇    下一篇

RNA干扰组蛋白去乙酰化酶4抑制骨肉瘤细胞诱导凋亡

肖前仁1,黄路2,张中卒3,张战民1,陈少卿1,舒勇1,曹凯1   

  1. 1. 南昌大学第一附属医院
    2. 江西省妇幼保健院
    3. 重庆医科大学附属永川医院
  • 收稿日期:2016-02-25 修回日期:2016-05-03 出版日期:2016-12-05 发布日期:2016-11-29
  • 通讯作者: 曹凯 E-mail:kaichaw@126.com
  • 基金资助:
    从表观遗传调控角度探讨miRNA-140对骨肉瘤恶性表型的抑制作用

RNA interfering histone deacetylase 4 induces apoptosis of osteosarcoma cells

  • Received:2016-02-25 Revised:2016-05-03 Online:2016-12-05 Published:2016-11-29

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(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HDAC4 mRNA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。

关键词: 骨肉瘤, RNA干扰, 组蛋白去乙酰化酶4

Abstract: Objective:To explore the functions of HDAC4 gene on the biological behaviorsof HOS and U2OS osteosarcoma cells lines whichinhibit HDAC4 gene expression using a small interfering RNA.Methods: The HOS and U2OS cells were transfected with a chemical synthesized small interfering RNA targeting HDAC4; The real time-PCR and Western blot assays were performed to detect the mRNA and protein expression levels of HDAC4 and its downstream genes HIF-1α, respectively; The CCK-8, flow cytometry (FCM), invasion assays were used to identifythe 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 U2OS cellsafter transfected with si-HDAC4 (P<0.01); in addition, the cell proliferation was inhibited and cell apoptosis was induced after suppression of HDAC4. (P<0.05); The cells of HOS and U2OS migrating through themembrane insi-con group and si-HDAC4 group were (146±34), (45±20)and (207±35)、(121±23), (P<0.05), respectively.Western blot showed that the expression of HDAC4 and HIF-1α was significantly reduced. Conclusion:HDAC4si-RNA can inhibit cell proliferation, invasion and induce cell apoptosis in human HOS and U2OSosteosarcoma cells.

Key words: Osteosarcoma, RNA interference, HDAC4