摘要
目的探讨锰型超氧化物歧化酶(MnSOD)基因转染对人骨肉瘤细胞的诱导凋亡作用。方法采用电穿孔方法将反义和正义MnSOD cDNA表达载体稳定转染OS-732骨肉瘤细胞,应用四唑蓝(MTT)比色法、流式细胞技术、DNA梯度实验观察MnSOD转染对OS-732细胞的诱导凋亡作用。结果转染了pHβA-SOD (+)载体的OS-732细胞(MnSOD-732)较对照空载细胞(Vector-732)生长明显受到抑制,生长曲线呈下降趋势(P＜0．05),说明细胞凋亡或死亡。转染了pHβA-SOD(-)载体的OS-732细胞(MnSOD AS-732)生长曲线呈上升趋势(P＜0．05),说明MnSOD低表达可促进骨肉瘤细胞生长。Vector-732和MnSOD AS-732培养24h后未出现Ap峰,MnSOD-732培养24h后G_1峰前出现Ap峰,肿瘤细胞凋亡率为13．26%。细胞周期分析显示, MnSOD-732较Vector-732或MnSOD AS-732的S期比例明显增加,G2～M期比例明显减少。MnSOD-732可见DNA梯度条带,Vector-732和MnSOD AS-732均未见明显梯状条带。结论转染正义MnSOD质粒可诱导OS-732细胞凋亡,MnSOD有望成为新的骨肉瘤细胞抑制剂。
Objective To appraise the apoptosis induction effect of maganese superoxide dismutase （MnSOD） gene transfection in human osteosarcoma cells. Methods We transfected sense and antisense MnSOD cDNA carrier PHβA-SOD （-）/primA-SOD （＋） or mock vector into OS-732 osteosarcoma cells by electroporation. Thiazolyl blue （MTT） test, flow cytometry （FCM） and agarose electrophoresis of DNA ladder were used to detect the proliferation activities and apoptosis rate. Results Compared with the OS-732 cells transfected with vector, the cells transfected with MnSOD（＋） had growth inhibition, the hypodiploid cusp appeared before G1 cusp by FCM and the DNA ladders through gel electrophoresis. On the contrary, the cells transfected with MnSOD （- ） showed enhancement of tumor proliferation without apoptosis characteristics. Conclusions MnSOD transfection can inhibit the growth and induce apoptosis of OS-732 cells. Thus MnSOD might be a promising growth inhibitor for osteosarcoma cells. （Shanghai Med J, 2007, 30： 442-445）
出处
《上海医学》
CAS
CSCD
北大核心
2007年第6期442-445,共4页
Shanghai Medical Journal
关键词
骨肉瘤
凋亡
基因转染
超氧化物歧化酶
Osteosarcoma
Apoptosis
Gene transfection
Superoxide dismutase
