基于CXCL5/FOXD1/VEGF信号通路探讨华蟾素对HepG2细胞的影响  

Effect of Cinobufacini on HepG2 cells based on CXCL5/FOXD1/VEGF pathway

作　　者：冉小柯 刘旭东[2] 庞华珍 谭伟强 吴铁雄 潘兆权 袁媛 娄鑫凤 RAN Xiao-ke;LIU Xu-dong;PANG Hua-zhen;TAN Wei-qiang;WU Tie-xiong;PAN Zhao-quan;YUAN Yuan;LOU Xin-feng(Graduate School,Guangxi University of Chinese Medicine,Nanning 530000,China;Dept of Hepatology,Ruikang Hospital Affiliated to Guangxi University of Chinese Medicine,Nanning 530000,China)

机构地区：[1]广西中医药大学研究生院,广西南宁530000 [2]广西中医药大学附属瑞康医院肝病科,广西南宁530000

出　　处：《中国药理学通报》2024年第12期2361-2368,共8页Chinese Pharmacological Bulletin

基　　金：广西岐黄学者建设项目;广西中医药大学岐黄工程高层次人才培育项目(No 2021007);广西中医药大学2023年研究生教育创新计划项目(No YCBXJ2023032)。

摘　　要：目的 探讨华蟾素对H ep G2细胞增殖、侵袭和凋亡的影响及其作用机制。方法 采用CCK-8法检测华蟾素对HepG2细胞增殖的影响;Transwell实验检测华蟾素对HepG2细胞侵袭的影响;流式细胞术检测华蟾素干预后HepG2细胞的凋亡比率;酶联免疫法检测HepG2细胞培养液中VEGF表达情况;qRT-PCR和Western blot检测华蟾素对CXCL5/FOXD1/VEGF信号通路中相关mRNA和蛋白表达水平的影响;使用免疫共沉淀检测CXCL5与FOXD1的结合情况。结果与对照组相比,随着华蟾素药物浓度的增加,HepG2细胞活力逐渐降低(P<0.05),并且华蟾素明显抑制HepG2细胞侵袭(P<0.05)、促进细胞凋亡(P<0.01),华蟾素能抑制CXCL5/FOXD1/VEGF信号通路中相关mRNA和蛋白的表达(P<0.05)。FOXD1是CXCL5的结合位点,CXCL5过表达后HepG2细胞出现增殖(P<0.05)和VEGF分泌增多(P<0.05),FOXD1和VEGF的表达增加(P<0.05);华蟾素干预后抑制肝癌细胞增殖和侵袭(P<0.05),促肝癌细胞凋亡(P<0.05),减少HepG2细胞分泌VEGF(P<0.05),降低HepG2细胞中CXCL5与FOXD1的表达(P<0.05),但与未过表达的华蟾素组相比其抑制细胞活性能力减弱(P<0.05),抑制CXCL5、FOXD1与VEGF表达的能力减弱(P<0.05)。结论 华蟾素可能通过调控CXCL5/FOXD1/VEGF信号通路抑制HepG2细胞增殖和侵袭,促进HepG2细胞凋亡。Aim To investigate the impact of Cinobu-facini on the proliferation,invasion,and apoptosis of HepG2 cells and the underlying mechanism.Methods The proliferation of HepG2 cells was assessed using the CCK-8 method following treatment with Cinobufacini.The invasion capability of HepG2 cells was evaluated through Transwell assay after exposure to Cinobufacini.The apoptosis rates of HepG2 cells post Cinobufacini intervention were measured using flow cytometry,and the expression levels of VEGF in the culture medium of HepG2 cells were determined using enzyme-linked immunoassay.Furthermore,qRT-PCR and Western blot analyses were conducted to assess the impact of Cinobufacini on mRNA and protein expression levels related to the CXCL5/FOXD1/VEGF pathway.The interaction between CXCL5 and FOXD1 was investigated via co-immunoprecipitation.Results Cinobufacini treatment led to a gradual decrease in HepG2 cell viability in a dose-dependent manner compared to the control group(P<0.05).Moreover,Cinobufacini significantly suppressed HepG2 cell invasion(P<0.05)while enhancing cell apoptosis(P<0.05).Notably,Cinobufacini exhibited inhibitory effects on the CXCL5/FOXD1/VEGF pathway,as evidenced by reduced expression of related mRNA and proteins(P<0.05).FOXD1 was identified as the binding site of CXCL5.Overexpression of CXCL5 resulted in increased proliferation and VEGF secretion by HepG2 cells(P<0.05),and increased expression of FOXD1 and VEGF(P<0.05).However,Cinobufacini intervention effectively inhibited liver cancer cell proliferation and invasion(P<0.05),promoted apoptosis(P<0.05),reduced VEGF secretion by HepG2 cells(P<0.05),and downregulated the expression of CXCL5 and FOXD1 in HepG2 cells(P<0.05);but compared with the unexpressed group of Cinobufacini,its ability to inhibit cell activity was weakened(P<0.05),and its ability to inhibit the expression of CXCL5,FOXD1,and VEGF was weakened(P<0.05).Conclusion Cinobufacini may inhibit HepG2 cell proliferation and invasion and promote HepG2 cell apoptosis by regulating the CXCL5/FOXD1

关 键 词：华蟾素 HEPG2细胞 CXCL5 FOXD1 VEGF 凋亡 

分 类 号：R284.1[医药卫生—中药学] R329.25[医药卫生—中医学] R329.28R392.12R735.7