论文部分内容阅读
[目的]探讨反义Snail转录因子对人骨肉瘤细胞株MG-63顺铂化疗敏感性的影响。[方法]用脂质体转染法将高转移性骨肉瘤细胞株MG-63分别转染反义Snail质粒(pGenesil-snailAS)和空载体质粒(pGenesil)。MG-63组(对照组)、MG-63/pGenesil组和MG-63/pGenesil-snailAS组细胞分别与浓度为0、0.5、1、5、10、15、20μg/ml顺铂作用72h。MTT法测定各组细胞的存活率并计算IC50,FCM检测细胞凋亡,Westernblot检测各组细胞Snail蛋白表达。[结果]细胞稳定转染pGenesil-snailAS后,细胞内Snail蛋白表达明显被抑制。MG-63、MG-63/pGenesil和MG-63/pGenesil-snailAS组细胞的顺铂IC50分别为14.0±1.8、12.6±1.6和2.8±0.5μg/ml,MG-63/pGenesil-snailAS组细胞对顺铂作用的敏感性增加了5倍。顺铂对MG-63/pGenesil-snailAS组细胞所诱导的凋亡比MG-63组和MG-63/pGenesil组显著。[结论]反义Snail转录因子能诱导人骨肉瘤细胞株MG-63凋亡并抑制其生长,增加其对顺铂化疗敏感性。
[Objective] To investigate the effect of antisense Snail transcription factor on chemosensitivity of human osteosarcoma cell line MG-63. [Method] The highly metastatic osteosarcoma MG-63 cells were transfected with pGenesil-snailAS and pGenesil respectively by lipofectamine. The cells in MG-63 group (control group), MG-63 / pGenesil group and MG-63 / pGenesil-snailAS group were treated with cisplatin at concentrations of 0, 0.5, 1, 5, 10, 15 and 20μg / ml respectively for 72h. Cell viability was measured by MTT assay and IC50 was calculated. FCM was used to detect cell apoptosis. Snail protein expression was detected by Western blot. [Result] After the cells were stably transfected with pGenesil-snail AS, the expression of Snail protein was obviously inhibited. The IC50 of cisplatin in MG-63, MG-63 / pGenesil and MG-63 / pGenesil-snailAS groups were 14.0 ± 1.8,12.6 ± 1.6 and 2.8 ± 0.5μg / ml, The sensitivity of cisplatin increased 5-fold. Cisplatin induced apoptosis in MG-63 / pGenesil-snail AS cells more significantly than MG-63 and MG-63 / pGenesil groups. [Conclusion] Antisense Snail transcription factor can induce the apoptosis of human osteosarcoma cell line MG-63 and inhibit its growth and increase its sensitivity to cisplatin chemotherapy.