香菇菌C91-3凋亡相关蛋白24414对人肺癌细胞A549凋亡作用及其机制的研究

目的:研究香菇菌C91-3凋亡相关蛋白24414对人肺癌细胞A549的凋亡诱导作用,探讨其诱导肿瘤细胞凋亡的机制。方法:将用毕赤酵母GS115体系进行诱导表达并已鉴定的香菇菌C91-3凋亡相关蛋白24414作用于人肺癌细胞A549,用MTT法测定目的蛋白的抑瘤率,用流式细胞术检测目的蛋白对肿瘤细胞的诱导凋亡作用,用ELISA法测定目的蛋白对肿瘤细胞Caspase-3活性的影响,同时用MTT法检测目的蛋白对正常鸡胚成纤维细胞的毒性作用。结果:流式细胞术与MTT法检测结果显示,终浓度为7.5、15和30μg/mL的此蛋白对A549细胞凋亡作用与对照组比较差异有统计学意义。ELISA法检测结果显示以上浓度的此蛋白对A549细胞Caspase-3活性影响与对照组相比较差异也有显著性意义(7.5μg/mL组vs对照组:t=8.787,P=0.000;15μg/mL组vs对照组:t=13.429,P=0.000;30μg/mL组vs对照组:t=36.908,P=0.000)。同时此蛋白对正常鸡胚成纤维细胞无毒性作用。结论:香菇菌C91-3凋亡相关蛋白24414在对正常鸡胚成纤维细胞无毒性作用的同时对人肺癌细胞A549具有诱导凋亡作用,其凋亡途径可能通过Caspase-3通路。

Apoptosis mechanism of Lentinus edodes C91-3 apoptosis protein 24414 on human lung cancer cell line A549

OBJECTIVE:To study the apoptosis function of the characterized protein(Lentinus edodes C91-3 apoptosis protein 24414) on human lung cancer cell line A549.METHODS:The interest protein was indueed to be expressed in Pichia pastoris GS115.Co-cultured the characterized protein with A549 cell was co-cultured and detected with flow cytometry to study the apoptosis function,with MTT method to study the inhibited rate,and with ELISA method to study the level of Caspase-3.At the same time,the toxic effect of the characterized protein was detected with MTT method by co-culturing the characterized protein with chick embryo fibroblast.RESULTS: The flow cytometry and MTT showed that there was a great difference between the interest protein groups(7.5,15 and 30 μg/mL) and control group according to the apoptosis function.The difference of Caspase-3 was also significant between the interest protein groups and control group(7.5 μg/mL group vs control group:t=8.787,P=0.000;15 μg/mL group vs control group:t=13.429,P=0.000;30 μg/mL group vs control group:t=36.908,P=0.000),While,there was no toxic effect of the characterized protein on chick embryo fibroblast.CONCLUSIONS: Lentinus edodes C91-3 apoptosis protein 24414 has a kind of apoptosis function on human lung cancer cell line A549 while has no toxic effect on normal cell.The fuction may be worked throug Caspase-3 pathway.