Survivin反义寡核苷酸对骨肉瘤细胞凋亡的影响

Antisense oligonucleotide targeting survivin gene induces cell apoptosis of osteosarcoma

目的:采用survivin反义寡核苷酸(ASODN)封闭骨肉瘤细胞survivin的作用,观察其对骨肉瘤细胞系MG63增殖和凋亡的影响。方法:以脂质体为载体,介导survivin反义寡核苷酸转染人骨肉瘤细胞系MG63,用MTT法测定细胞增殖;流式细胞仪检测转染survivin反义寡核苷酸后细胞凋亡率;Westernblot及半定量RTPCR检测转染survivin反义寡核苷酸后的骨肉瘤细胞survivin蛋白及mRNA表达。结果:脂质体介导survivin反义寡核苷酸转染骨肉瘤细胞系MG63后,细胞增殖明显受到抑制,细胞凋亡率明显增加,骨肉瘤细胞survivin蛋白及mRNA表达均明显降低。结论:脂质体介导转染survivin反义寡核苷酸可以抑制细胞增殖,有效降低细胞内survivin基因的表达,促进细胞凋亡。

Objective： The aim of this study was to investigate the effect of antisense oligonucleotide （ASODN） targeting survivin on proliferation and apoptosis of osteosarcoma cells line MG-63. Methods：Survivin ASODN was designed and transfected into MG-63 cells mediated by liposome carrier. Cultured cells were divided into 5 groups： control group, lipofection group, sense oligonucleotide （SODN） group, mismatch oligonucleotide （MODN） group, and ASODN group. Proliferation of ovarian cells was detected by methyl thiazolyl tetrazolium （MTT） assay; apoptotic rate was determined by flow cytometry; survivin protein and mRNA expression were measured by western blot and reversed transcriptase polymerase chain reaction, respectively. Results： The proliferation of MG-63 cells was inhibited notably after ASODN transfection and the apoptosis rate of ASODN groups was significantly higher than that of other groups. Survivin protein and mRNA expression in ASODN group were significantly decreased compared with that in the control groups. Conclusions： Liposome-mediated survivin ASODN transfection significantly inhibits the proliferation of osteosarcoma MG-63 cells, reduces the expression of survivin genes,and induces cell apoptosis.