化癥散积方联合顺铂对HepG2肝癌细胞的增殖和迁移的影响  

Effects of Huazheng Sanji Fang in combination with cisplatin on the proliferation and migration of liver cancer cells HepG2

作　　者：袁璐 刘扬扬[2] 徐晓浩 王珊珊[3] 孟香彤 刘铁军[1,2] 冷炎[1,2] 熊壮[1,2] YUAN Lu;LIU Yangyang;XU Xiaohao;WANG Shanshan;MENG Xiangtong;LIU Tiejun;LENG Yan;XIONG Zhuang(Changchun University of Chinese Medicine,Changchun 130117,China;Affiliated Hospital of Changchun University of Chinese Medicine,Changchun 130021,China;The Second Affiliated Hospital of Shandong University of Traditional Chinese Medicine,Jinan 250001,China)

机构地区：[1]长春中医药大学,长春130117 [2]长春中医药大学附属医院,长春130021 [3]山东中医药大学第二附属医院,济南250001

出　　处：《吉林中医药》2024年第4期461-466,共6页Jilin Journal of Chinese Medicine

基　　金：国家自然科学基金项目(81804007);吉林省自然科学基金项目(20200201590JC)。

摘　　要：目的通过单独使用化癥散积方(HZSJ)或与顺铂(cisplatin,DDP)联合应用干预HepG2肝癌细胞的增殖和迁移,以探讨其治疗肝癌的潜在机制。方法将HepG2肝癌细胞分为空白对照组、缺氧模型组、DDP组、HZSJ组、HZSJ+DDP组,采用CoCl2法诱导缺氧环境。通过MTT法、流式细胞术和细胞划痕试验验证HZSJ联合DDP对HepG2肝癌细胞增殖、凋亡和迁移的影响。采用酶联免疫吸附测定(ELISA)检测凋亡相关因子;网络药理学进行靶点预测;蛋白免疫印迹法(Western blot)检测HZSJ联合DDP对HepG2肝癌细胞中低氧诱导因子(HIF-1α)和血管内皮生长因子(VEGF)表达的影响。结果HZSJ和DDP可以显著抑制HepG2肝癌细胞的增殖,并且HZSJ与DDP具有协同作用,降低DDP的有效浓度;当低浓度DDP和HZSJ联合应用时,HepG2肝癌细胞的早期和晚期凋亡数量显著增加,Bax/Bcl-2的比值显著增加,Cleaved-Casp-3的表达水平也显著增加;低浓度DDP不影响HepG2肝癌细胞的迁移,而高浓度HZSJ可抑制HepG2的迁移,当两者联合应用时能够显著降低HepG2肝癌细胞的迁移率;基于STRING数据库构建HZSJ潜在靶点网络图,确定了HIF信号通路是HepG2肝癌细胞增殖和迁移的主要调节通路;Western blot结果表明,HZSJ和低浓度DDP联合应用时,HIF-1α和VEGF的表达水平显著降低。结论HZSJ可以帮助DDP抑制HepG2肝癌细胞的增殖和迁移,两者协同促进HepG2肝癌细胞凋亡,并调节凋亡相关蛋白Bax、Bcl-2、Cleaved-Casp-3以及迁移相关蛋白HIF-1α和VEGF。Objective To explore its potential mechanisms of Huazheng Sanji Fang(HZSJ)on the treatment of liver cancer by means of intervening in the proliferation and migration of liver cancer cells HepG2 with the Chinese medicine alone or in combination with cisplatin(DDP).Methods Liver cancer cells HepG2 were divided into a blank control group,a hypoxic model group,a DDP group,an HZSJ group,an HZSJ+DDP group,and the hypoxic environment was induced by the CoCl2 method.The effects of HZSJ combined with cisplatin on the liver cancer cells HepG2 proliferation,migration and apoptosis were verifi ed by the MTT method,flow cytometry assay and cell scratch test.Enzyme-linked immunosorbent assay(ELISA)was used to detect the apoptosis-related factors.The effects of HZSJ and cisplatin on the expression of hypoxia-inducible factor-1α(HIF-1α)and vascular endothelial growth factor(VEGF)in HepG2 cells were detected by Western blot.Results HZSJ and cisplatin significantly inhibited the proliferation of HepG2 cells,and they had a synergistic effect,reducing the effective concentration of cisplatin.The number of early and late apoptosis of HepG2 cells was significantly increased under the mixed trial of low concentration of cisplatin and HZSJ.Simultaneously,the ratio of Bax/Bcl-2 levels was also increased significantly,and the expression level of Cleaved-Casp-3 was significantly increased as well.Low-dose cisplatin did not aff ect the migration of HepG2 cells,while high-dose HZSJ inhibited its migration,and when mixed,the two drugs at low concentration had a significant inhibitory effect.Based on the STRING database,a potential target network diagram of HZSJ was constructed,and the HIF signaling pathway was identifi ed as the main regulatory pathway for the proliferation and migration of HepG2 cells.Western blot results showed that the combination of HZSJ and low concentration DDP significantly reduced the expression levels of HIF-1αand VEGF.Conclusion HZSJ can assist cisplatin in inhibiting the migration and proliferation of HepG2 cell

关 键 词：中医药 细胞增殖 细胞凋亡 细胞迁移 肝癌 

分 类 号：R735.7[医药卫生—肿瘤]