基于EGFR/AKT和JAK2/STAT3通路研究α-常春藤皂苷单独或与顺铂联用对非小细胞肺癌细胞增殖与凋亡的影响  

Effects of α-hederin alone or in combination with cisplatin on the proliferation and apoptosis of non-small cell lung cancer cells based on EGFR/AKT and JAK2/STAT3 pathways

作　　者：朱志明[1] 王苏美 唐青 王晰 万信良 莫瀚丹 贾璐瑜 俞晓燕 周绮纯 ZHU Zhiming;WANG Sumei;TANG Qing;WANG Xi;WAN Xinliang;MO Handan;JIA Luyu;YU Xiaoyan;ZHOU Qichun(Department of Pharmacy,the Second Clinical College of Guangzhou University of Chinese Medicine,Guangzhou 510120,Guangdong,China;Clinical and Basic Research Team of TCM Prevention and Treatment of Non-Small Cell Lung Cancer,the Second Clinical College of Guangzhou University of Chinese Medicine,Guangzhou 510120,Guangdong,China)

机构地区：[1]广州中医药大学第二临床医学院药剂科,广东广州510120 [2]广州中医药大学第二临床医学院中医药防治非小细胞肺癌临床与基础研究团队,广东广州510120

出　　处：《中国肿瘤生物治疗杂志》2024年第4期333-341,共9页Chinese Journal of Cancer Biotherapy

基　　金：广东省中医院拔尖人才科研专项(No.BJ2022KY13);广东省中医药局面上项目(No.20222085,No.20231094)。

摘　　要：目的:探讨α-常春藤皂苷(α-Hed)诱导非小细胞肺癌(NSCLC)细胞凋亡的作用靶点及其潜在机制,明确α-Hed与顺铂(DDP)联用后对相应的靶点蛋白表达的影响。方法:采用CCK-8法检测不同浓度α-Hed处理后NSCLC细胞A549、H1299和PC-9的存活率,采用AnnexinⅤ-FITC/PI染色流式细胞术检测细胞凋亡率,采用WB法检测细胞中C-caspase-3和Bcl-2蛋白的表达。通过网络药理学相关方法筛选α-Hed的潜在靶点,利用分子对接法分析其结合效果,WB法检测靶点蛋白的表达。通过CCK-8法、细胞集落形成实验和WB法检测α-Hed与DDP联用对NSCLC细胞的抑制作用。结果:给药24和48 h后,10、15和20μmol/Lα-Hed可以显著抑制NSCLC细胞增殖活力(均P<0.01);与对照组相比,20μmol/Lα-Hed处理后细胞凋亡率显著升高(P<0.01);α-Hed可上调NSCLC细胞中C-caspase-3的表达(P<0.05),下调Bcl-2的表达(P<0.05)。网络药理学和分子对接筛选出结合亲和力小于-5 kcal/mol的靶点AKT1、STAT3、EGFR和JAK2。WB法检测结果显示,α-Hed处理后A549、H1299细胞中EGFR、p-AKT/AKT、p-STAT3/STAT3和JAK2蛋白的表达均明显下调(均P<0.05)。α-Hed与DDP联用后,更显著地抑制NSCLC细胞的增殖(P<0.01),进一步下调EGFR、p-AKT/AKT、p-STAT3/STAT3和JAK2蛋白的表达(P<0.05或P<0.01)。结论:α-Hed通过下调EGFR和JAK2的表达抑制STAT3和AKT的磷酸化,诱导NSCLC细胞凋亡,与DDP联用后其抑制效果增强,EGFR/AKT和JAK2/STAT3通路也进一步被抑制。Objective:To explore the targets and potential mechanism ofα-hederin(α-Hed)inducing cell apoptosis on non-small cell lung cancer(NSCLC),and to clarify the effects of α-Hed in combination with cisplatin(DDP)on the expression of target proteins.Methods:The viability of NSCLC cells(A549,H1299 and PC-9 cells)treated with different concentrations of α-Hed was detected by CCK-8 method.The apoptosis rate was determined by flow cytometry with AnnexinⅤ-FITC/PI staining.The expressions of cleaved caspase-3(C-caspase-3)and Bcl-2 proteins were detected by Western blot.The potential targets ofα-Hed were screened by network pharmacology,and their binding effect was analyzed by molecular docking method.Besides,Western blot was applied to detect target protein expression.The suppressive effect ofα-Hed in combination with DDP on NSCLC cells was detected by CCK-8 assay,colony formation assay and Western blot assay.Results:After 24 h and 48 h administration,α-Hed at 10,15 and 20μmol/L significantly inhibited the proliferation viability of NSCLC cells(all P<0.01).Compared with the control group,the apoptosis rate significantly increased after 20μmol/Lα-Hed treatment(P<0.01).C-caspase-3 expression in NSCLC cells was upregulated(P<0.05)and Bcl-2 expression was downregulated afterα-Hed administration.The targets of AKT1,STAT3,EGFR and JAK2 with the binding affinity less than-5 kcal/mol were screened out via network pharmacology and molecular docking.Western blot showed that the expressions of EGFR,p-AKT/AKT,p-STAT3/STAT3 and JAK2 proteins in A549 and H1299 cells were significantly downregulated after α-Hed treatment(all P<0.05).Furthermore,α-Hed in combination with DDP more significantly inhibited the proliferation of NSCLC cells(P<0.01)and downregulated the expression of EGFR,p-AKT/AKT,p-STAT3/STAT3 and JAK2 proteins(P<0.05 or P<0.01).Conclusion:α-Hed induces the apoptosis of NSCLC by down-regulating EGFR and JAK2 expressions,and inhibiting the phosphorylation of STAT3 and AKT.Especially,the inhibitory effect is enhanc

关 键 词：α-常春藤皂苷 非小细胞肺癌 增殖 凋亡 AKT1 STAT3 EGFR JAK2 

分 类 号：R734.2[医药卫生—肿瘤] R730.52[医药卫生—临床医学]