论文部分内容阅读
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。
Objective: To study the apoptosis-inducing effect of Lentinula edodes C91-3 apoptosis-associated protein 24414 on human lung cancer cell line A549 and to explore its mechanism of inducing tumor cell apoptosis. Methods: The human leukemia cell line A549 was induced by the Pichia pastoris GS115 system and identified as Lentinula edodes C91-3 apoptosis related protein 24414. The inhibition rate of the target protein was determined by MTT assay. The expression of the target protein was detected by flow cytometry The target protein was used to induce apoptosis of tumor cells. The effect of the target protein on the activity of Caspase-3 was determined by ELISA. Meanwhile, the toxicity of target protein on normal chicken embryo fibroblasts was detected by MTT assay. Results: The results of flow cytometry and MTT assay showed that the apoptosis of A549 cells at the final concentration of 7.5, 15 and 30 μg / mL was significantly different from that of the control group. The results of ELISA showed that the above concentration of this protein had significant difference on the activity of Caspase-3 in A549 cells compared with the control group (7.5μg / mL vs control group: t = 8.787, P = 0.000; 15μg / mL group vs control group: t = 13.429, P = 0.000; 30μg / mL vs control: t = 36.908, P = 0.000). At the same time, this protein has no toxic effect on normal chicken embryo fibroblasts. CONCLUSION: The apoptosis-related protein 24414 of Lentinus edodes can induce apoptosis in human lung cancer cell A549 while its non-toxic effect on normal chicken embryo fibroblasts. The apoptotic pathway may be through the Caspase-3 pathway.