摘要
目的研究caspase-3在肿瘤特异性凋亡基因诱导人黑色素瘤细胞A375凋亡中的作用。方法用含有apoptin基因的真核表达载体瞬间转染体外培养的人黑色瘤细胞A375;采用RT-PCR、DNA凝胶电泳、流式细胞术检测A375细胞的凋亡;以比色法检测caspase-3的相对活性。结果Ap-optin基因瞬间转染的A375细胞可出现典型的细胞凋亡所具有的DNA梯状带;流式细胞术发现实验组细胞凋亡率明显高于其他各组(P<0.01);转染后24h实验组caspase-3的活性开始升高,72h达高峰,明显高于其他各组(P<0.01)。结论Apoptin基因可通过激活caspase-3诱导人黑色素瘤细胞A375凋亡。
AIM： To study whether apoptin gene induces apoptosis in human melanoma cells A375 via the activation caspase-3. METHODS： The human melanoma cells A375 were transiently transfected with recombinant pcDNAA3 plasmid, which contained apoptin gene, and the apoptosis was measured by DNA agarose electrophoresis, RT-PCR and flow cytometry, caspase-3 relative activity was analyzed by colorimetric assay at various time. RESULTS： At 48 h after transfecUon the apoptin gene could induce apoptosis of human melanoma cells A375 in vitro and the mRNA of apoptin gene could be detected by RT-PCR. Caspase-3 activity began to rise at 24 h after transfection and reached the peak at 72 h. CONCLUSION： Apoptin gene can induce apoptosis in human melanoma cells A375 via active caspase-3 and the activity of caspase-3 is timedependent.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2006年第3期293-295,共3页
Chinese Journal of Cellular and Molecular Immunology
基金
辽宁省自然科学基金资助课题(No.20042046)
