基础医学与临床 ›› 2009, Vol. 29 ›› Issue (5): 527-530.

• 研究论文 • 上一篇    下一篇

Livin反义寡核苷酸对人HL60细胞增殖和凋亡的影响

周慷 兰箭   

  1. 重庆医科大学 第二临床学院 血液内科 重庆医科大学 第二临床学院 血液内科
  • 收稿日期:2008-06-23 修回日期:2008-10-23 出版日期:2009-05-25 发布日期:2009-05-25
  • 通讯作者: 兰箭

Effects on Livin antisense oligodeoxynucleotides of the proliferation and apoptosis of HL60 cells

Kang ZHOU, Jian LAN   

  1. Department of Hematology,the Second Affiliated Hospital of Chongqing Medical University Department of Hematology,the Second Affiliated Hospital of Chongqing Medical University
  • Received:2008-06-23 Revised:2008-10-23 Online:2009-05-25 Published:2009-05-25
  • Contact: Jian LAN

摘要: 目的 探讨Livin mRNA反义寡核苷酸(ASODN)对人白血病细胞(HL60)增殖及凋亡的影响。方法 用免疫组织化学检测Livin蛋白表达,设计合成特异性的Livin硫代磷酸ASODN及其对照错义寡核苷酸(MSODN),脂质体转染HL60细胞。用四甲基偶氮唑盐光吸收法(MTT)检测细胞增殖,反转录-聚合酶链反应(RT-PCR)检测Livin mRNA的表达,原位末端标记技术(TUNEL)、电镜检测细胞凋亡率和形态学改变。结果Livin ASODN在终浓度为600nmol/L作用HL60细胞48h时,能明显地抑制其增殖,降低Livin mRNA的表达,HL60细胞在形态学上出现明显的凋亡改变,细胞凋亡率显著增加 (P<0.01)。结论Livin ASODN可有效抑制HL60细胞的增殖,下调Livin基因表达,并促进HL60细胞凋亡,在诱导肿瘤细胞凋亡、抑制增殖中发挥重要作用。

关键词: 白血病, Livin基因, 反义寡核苷酸类, 细胞凋亡

Abstract: Objective To investigate effects of the proliferation and apoptosis of human leukemia ( HL60) cells by Livin antisense oligodeoxynucleotides (ASODN).Methods to detect expression of Livin protein on HL60 cells by immunohistochemistry. Specific phosphorothioate antisense oligodeoxynucleotides and missense oligodeoxynucleotides target Livin mRNA were synthesized and was transfected into HL60 cells following cationic liposome.The proliferation inhibitory rates of HL60 cells were assessed by MTT method. The expressions of Livin mRNA were detected by RT-PCR. Transmission electron microscope and TUNEL technology were used to detect the apoptosis level and morphologic changes. Results ASODN at a final concentration of 600 nmol/L could inhibit the HL60 cells proliferation and the expressions of Livin mRNA. The apoptosis rates detected by TUNEL was 38.48%±4.37%. cellar ultrastructure was markedly destroyed by Livin ASODN. There were significant differences compared with the control group (P<0.01).Conclusion Livin ASODN could suppress HL60 cell proliferation effectively, decline Livin gene expression, induce significant apoptosis of HL60, therefore Livin gene may become a new target for gene therapy of leukemia.

Key words: leukemia, Livin gene, Antisense oligodeoxynucleotides, Apoptosis