美洲大蠊多肽和顺铂联用对HepG2细胞凋亡及自噬的影响  

EFFECTS OF PERIPLANETA AMERICANA PEPTIDE COMBINED WITH CISPLATIN ON APOPTOSIS AND AUTOPHAGY OF HepG2 CELLS

作　　者：陈莉[1] 齐烨迪 李智 余丽双[1] CHEN Li;QI Ye-di;LI Zhi;YU Li-shuang(Pharmacy College,Fujian University of Traditional Chinese Medicine,Fuzhou,Fujian 350122,China)

机构地区：[1]福建中医药大学药学院,福建福州350122

出　　处：《井冈山大学学报（自然科学版）》2023年第4期69-74,共6页Journal of Jinggangshan University (Natural Science)

基　　金：国家自然科学青年基金项目(81903855)。

摘　　要：采用MTT法检测美洲大蠊多肽(PAP-3)与不同浓度的顺铂(DDP)单用或联合使用对人肝癌HepG2细胞增殖的影响;采用流式细胞术检测PAP-3、DDP单独或联合给药干预对HepG2细胞凋亡的影响;Western Blot检测PAP-3、DDP单独或联合给药后HepG2细胞内自噬相关蛋白p62、LC3、Beclin-1、Atg5及PI3K的表达。实验结果显示,与DDP单独给药相比,PAP-3与DDP联合给药对HepG2的细胞的抑制作用更强;PAP-3与DDP联合给药组中细胞的凋亡率与DDP和PAP-3单独给药组相比均有升高(P<0.05)。与对照组相比,DDP组中,p62蛋白水平降低,LC3II蛋白水平升高,LC3II/I的比例也升高,表现出细胞自噬流的活化;与DDP组相比,联合给药组中p62蛋白水平回升,LC3II蛋白水平和LC3II/I的比例均有回落。且联合给药组中PI3K、Atg5、Beclin-1等自噬相关蛋白的量均较DDP组减少,而凋亡相关的Caspase-3蛋白的表达量则较DDP组增加。据此推测,PAP-3和DDP的联用可以通过抑制HepG2细胞自噬相关蛋白的表达水平,抑制DDP带来的细胞自噬水平的升高,增加细胞对DDP的敏感性,从而诱导HepG2细胞凋亡。MTT assay was used to detect the effects of Periplaneta americana peptide(PAP-3,sequence IPMTPAGI-NH2)and cisplatin(DDP)alone or in combination on the proliferation of HepG2 cells.The effects of PAP-3 and DDP alone or in combination on apoptosis in HepG2 cells were detected by flow cytometry.Western blotting was used to detect the expression levels of autophagy associated proteins p62,LC3,Beclin-1,Atg5 and PI3K in HepG2 cells after drug administered in each group.Compared with DDP alone,the combined administration of PAP-3 and DDP had stronger cytostatic effects on HepG2 cells.The apoptosis rate of PAP-3 and DDP combined group was higher than that of DDP and PAP-3 alone group(P<0.05),and the trend was similar to that of DDP+3-MA group.Compared with the control group,the p62 protein level in DDP group was significantly decreased,the LC3 protein level and the LC3II/I ratio were increased,showing the activation of autophagy flow.In contrast,in the PAP-3+DDP combined administration group,the p62 protein level increased,while the level of LC3II protein and the ratio of LC3II/I both decreased.The amount of autophagy related proteins such as PI3K,ATG5 and Beclin-1 in the PAP-3+DDP combined administration group was lower than that in the DDP group,while the expression of apoptosis related caspase-3 protein was higher than that in the DDP group.The combination of PAP-3 and DDP could increase the sensitivity of the cells to DDP and induce apoptosis of HepG2 cells by inhibiting the expression of autophagy-related proteins and inhibiting the increase of autophagy level caused by DDP.

关 键 词：美洲大蠊多肽 肝癌 凋亡 自噬 顺铂 

分 类 号：Q255[生物学—细胞生物学]