Livin反义寡核苷酸对人HL60细胞增殖和凋亡的影响

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

目的探讨LivinmRNA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L ivin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LivinmRNA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LivinASODN在终浓度为600 nmol/L作用HL60细胞48 h时,能明显地抑制其增殖,降低LivinmRNA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LivinASODN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。

Objective To investigate effects of Livin antisense oligodeoxynucleotides （ASODN） on the proliferation and apoptosis of human leukemia （HL60） cells. Methods Livin protein on HL60 cells was examined by immunohistochemistry. Specific phosphorothioate antisense oligodeoxynucleotides and missense oligodeoxynucleotides target Livin mRNA were synthesized and transfected into HL60 cells following cationic liposome. The proliferation inhibition of HL60 cells was assessed by MTT. The expression of Livin mRNA was detected by RT-PCR. Transmission electron microscope and TUNEL technology were used to detect the apoptosis and morphologic change. Results ASODN of 600 nmol/L inhibited the HL60 cell proliferation and the ,expressions of Livin mRNA. The percentage of apoptosis detected by TUNEL was 38.48%±4. 37%. cellar uhrastructure was markedly destroyed by Livin ASODN. A significant difference was found when compared with the control group （P 〈 0. 01 ）. Conclusion Livin ASODN may suppress HL60 cell proliferation effectively, decrease Livin gene expression and induce significant apoptosis of HL60. Livin gene is a potential target for gene therapy of leukemia.