Survivin反义寡核苷酸对骨肉瘤MG-63细胞系增殖和凋亡的影响

Effects of Survivin antisense oligodeoxynucleotide on proliferation and apoptosis in osteosarcoma cell line MG-63

目的 观察Survivin基因反义寡核苷酸对骨肉瘤细胞增殖和凋亡的影响。方法 靶向Survivin基因的反义寡核苷酸经脂质体介导转染骨肉瘤MG 63细胞系,倒置显微镜观察转染后细胞形态学变化、半定量RT PCR检测Survivin基因mRNA的表达,噻唑蓝 (MTT)法检测细胞的增殖情况,吖啶橙荧光染色法观察细胞凋亡情况。结果 转染反义寡核苷酸后,MG 63细胞中Survivin基因mRNA水平表达显著下降;细胞增殖受到显著抑制,抑制率达 61%;倒置显微镜显示细胞变圆,漂浮细胞增多;吖啶橙染色显示转染组细胞出现明显核碎裂等凋亡改变。结论 Survivin基因的反义寡核苷酸可以显著抑制骨肉瘤细胞增殖,促进细胞凋亡。

Objective To investigate the effects of Survivin antisense oligodeoxynucleotide (ODN) on proliferation and apoptosis in osteosarcoma cell line MG-63.Methods The antisense oligodeoxynucleotide targeting Survivin gene was transfected into MG-63 cells by liposomal reagent. Morphological change of tumor cells was observed under invert microscope. The expression of Survivin gene was detected by semi-quantitative reverse transcription PCR. The proliferation of cells was determined by MTT. Cell apoptosis was detected by acridine orange staining.Results The expression of surviving mRNA was down-regulated by surviving antisense ODN. The proliferation of tumor cells was significantly inhibited, and the growth rate in transfected cells was down to 61% of normal cells. The number of drift cells was increased after transfection. Apoptotic changes including nuclear fragmentation in transfected cells were observed by acridine orange staining.Conclusions Antisense ODN targeting Survivi gene could significantly inhibit the proliferation of MG-63 cells and increase the cell apoptosis.