地鳖纤溶活性蛋白含药血清对人肺癌A549细胞凋亡及凋亡相关表达的影响

Effect of protein of Eupolyphaga sinensis Walker containing Serum on induction of apoptosis and regulation of apoptosis related genes in A549 cell line

目的研究地鳖纤溶活性蛋白(EFP)含药血清对人肺癌A549细胞凋亡的影响及凋亡相关基因的表达。方法制备不同浓度EFP小鼠含药血清;采用MTT法检测EFP对A549细胞的增殖抑制效果;采用Annexin V-FITC/PI双荧光染色观察细胞凋亡情况;应用流式细胞术检测EFP对A549细胞周期的影响;采用免疫组化法检测凋亡相关基因Bax表达水平的改变。结果 EFP含药血清各组对A549细胞有明显抑制作用;EFP使A549细胞发生细胞凋亡,细胞周期发生改变;随着药物浓度的增加,Bax表达逐渐增强,Bcl-2表达逐渐减弱。结论 EFP有较强的体外抗肿瘤活性;EFP可能通过调控Bcl-2,Bax基因表达而诱导凋亡。

Objective To investigate whether protein of Eupolyphaga sinensis walker(EFP) containing Serum can induce apoptosis of A549 cell line and elucidate the regulation of EFP on Bcl-2 family genes. Methods EFP containing Serum was prepared.MTT assay was used to measured the anti-proliferative effect of EFP,and apoptosis was observed by Annexin V-FITC/PI double fluorescenec dyeing;Cell cycle and apoptosis of A549 cells were analyzed by flow cytometry(FCM).Alteration of protein of Bax genes was detected by immunohistochemical techniques. Results Medicated Serum had obvious anti-proliferation activity,A549 cells appeared notable apoptosis after incubated with EFP,and the cell cycle was changed by EFP.The Bax protein kept increasing during EFP treatment,while the Bcl-2 protein kept decreasing. Conclusion The protein of Eupolyphaga has an obvious inhibitory effect on tumor cell.The anti-tumor response of EFP in A549 cells is through down-regulation of protein expression of Bcl-2 genes and up-regulation of protein expression of Bax genes.