扶正抗癌方通过逆转HepG2细胞EMT抗肝癌的实验研究
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摘要
目的:探讨扶正抗癌方对TGF-β1诱导的HepG2细胞EMT的影响及扶正抗癌方抑制肝癌侵袭与转移的机制。方法:建立肝癌HepG2细胞EMT模型并将其分为模型组、顺铂组、扶正抗癌方组、联合用药组,分别加入扶正抗癌方与顺铂含药血清,通过黏附实验、划痕实验、Transwell实验测定细胞的侵袭及转移能力,并检测EMT相关蛋白的表达。结果:各给药组细胞的黏附率、划痕的愈合率、转移的HepG2细胞数均低于模型组(P<0.05或P<0.01),联合用药组转移的HepG2细胞数低于顺铂组(P<0.01);各给药组HepG2细胞中Vimentin表达低于模型组,而E-cadherin表达高于模型组(P<0.05或P<0.01),联合用药组HepG2细胞中Vimentin表达低于顺铂组,而E-cadherin表达高于顺铂组(P<0.01)。结论:扶正抗癌方通过上调上皮标志蛋白E-cadherin的表达,下调间质细胞标记物Vimentin的表达,降低HepG2细胞运动和侵袭性,逆转其EMT进程。
Objective: To explore the effect of Fuzheng Kang'ai Formula on TGF-β1-induced EMT of HepG2 cells and the mechanism of Fuzheng Kang'ai Formula on inhibiting the invasion and metastasis of hepatocellular carcinoma. Methods: Establised HepG2 cells EMT model and divided them into model group, Fuzheng Kang'ai Formula group, DDP group, Combination group, respectively joined Fuzheng Kang'ai Formula Recipe and Cisplatin-containing serum; We used cell-matrix adhesion assay, scratch test, transwell invasion and migration assay to determine the ability of HepG2 cells to adhere, invade and migrate; Western blot was used to detect the expression of EMT-related proteins. Results: The adhesion rate of the cells, healing rate of the scratches and transferred HepG2 cells in each medication group were lower than those in the model group(P<0.05 or P<0.01). The number of HepG2 cells transferred from Combination group was lower than that in the DDP group(P<0.01); The medication groups showed lower expression of Vimentin in HepG2 cells andhigher expression of E-cadherin than model group(P <0.05 or P <0.01). The expression of Vimentin was statistically significant in HepG2 cells. The combination group showed lower expression of Vimentin andhigher expression of E-cadherin than that of the DDP group(P<0.01). Conclusion: Fuzheng Kang'ai Formula can increase the expression of Ecadherin in the HepG2 cells, down-regulate the expression of the stromal cell marker Vimentin, reduce the single cell movement and invasiveness, and reverse the EMT process.
Objective: To explore the effect of Fuzheng Kang'ai Formula on TGF-β1-induced EMT of HepG2 cells and the mechanism of Fuzheng Kang'ai Formula on inhibiting the invasion and metastasis of hepatocellular carcinoma. Methods: Establised HepG2 cells EMT model and divided them into model group, Fuzheng Kang'ai Formula group, DDP group, Combination group, respectively joined Fuzheng Kang'ai Formula Recipe and Cisplatin-containing serum; We used cell-matrix adhesion assay, scratch test, transwell invasion and migration assay to determine the ability of HepG2 cells to adhere, invade and migrate; Western blot was used to detect the expression of EMT-related proteins. Results: The adhesion rate of the cells, healing rate of the scratches and transferred HepG2 cells in each medication group were lower than those in the model group(P<0.05 or P<0.01). The number of HepG2 cells transferred from Combination group was lower than that in the DDP group(P<0.01); The medication groups showed lower expression of Vimentin in HepG2 cells andhigher expression of E-cadherin than model group(P <0.05 or P <0.01). The expression of Vimentin was statistically significant in HepG2 cells. The combination group showed lower expression of Vimentin andhigher expression of E-cadherin than that of the DDP group(P<0.01). Conclusion: Fuzheng Kang'ai Formula can increase the expression of Ecadherin in the HepG2 cells, down-regulate the expression of the stromal cell marker Vimentin, reduce the single cell movement and invasiveness, and reverse the EMT process.
引文
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[15]肖跃红.原发性肝癌的中医病机及防治探讨[J].中国中医基础医学杂志,2013,19(6):617-648.
[16]秦英刚,花宝金.花宝金治疗肝癌经验[J].北京中医药,2013,32(1):33-34.
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[18]刘俊保,刘延庆.原发性肝癌的中医药治疗研究述评[J].中医学报,2013,28(1):11-13.
[19]李川,吕文良,孙桂芝.孙桂芝教授益气活血解毒散结法治疗肝癌学术思想[J].长春中医药大学学报,2012,28(6):1002-1003.
[20]郝丽君.浅谈陈光伟教授扶正抗癌法治疗肿瘤经验[J].中国民族民间医药,2013,22(11):127.
[21]杨柳青,陈光伟,陈建婷.扶正抗癌汤对肝癌小鼠突变型p53基因和巨噬细胞CD_(68)表达的影响[J].陕西中医,2011,32(10):1426-1427.
[22]方瑜,杨柳青,陈光伟.扶正抗癌方对小鼠肝癌细胞转移影响的实验研究[J].陕西中医,2013,34(9):1253-1255.
[23]方瑜,杨柳青,陈光伟.扶正抗癌方对H_(22)移植瘤小鼠凋亡和端粒酶表达影响的实验研究[J].陕西中医,2013,34(10):1423-1425.
[24]方瑜,杨柳青,王松岩,等.扶正抗癌方对H_(22)细胞STAT3通路的影响[J].陕西中医,2014,35(5):618-619.
[25]方瑜,张淑珍,陈光伟.扶正抗癌方抑制HepG2细胞诱导EMT机理探讨[J].陕西中医药大学学报,2016,39(6):121-124.
[26]彭凯丽,陈宏辉.上皮间质细胞转化在肿瘤侵袭转移及化疗耐药中的作用[J].实用医药杂志,2018,35(1):85-87.
[27]迟笑怡,胡凯文,周天.盐酸川芎嗪对小鼠Lewis肺癌细胞转移及上皮-间质转化的影响[J].中医药导报,2018,24(11):9-15,20.
[2]黄赞松,仇仪英,周喜汉.原发性肝癌现代医学治疗的研究进展[J].医学综述,2012,18(24):4169-4172.
[3]Yang T,Zhang J,Lu J H,et al.A new staging system for resectablehepatocellular carcinoma:comparison with six existing staging sys-tems in a large Chinese cohort[J].J Cancer Res Clin Oncol,2011,137(5):739-750.
[4]唐旭,胡志芳.抗肝癌治疗诱导的肝癌细胞上皮-间质转化研究进展[J].转化医学杂志,2013,2(4):109-112.
[5]Waldmeier L,Meyerschaller N,Diepenbruck M,et al.Py2Tmurine breast cancer cells,a versatile model of TGFβ-inducedEMT in vitro and in vivo[J].PLoS One,2012,7(11):e48651.
[6]吴艳林,刘润田.人参皂苷Rh2对人肝癌细胞HepG2/ADM耐药逆转作用及其机制研究[J].医学研究生学报,2017,30(5):476-480.
[7]齐明华.上皮间质转化及相关信号通路在原发性肝细胞癌转移中的研究进展[J].世界华人消化杂志,2012,20(11):953-958.
[8]吴冬,柯爱武,郭传勇.上皮间质转化在肝纤维化和肝细胞癌发病中的作用研究进展[J].实用肝脏病杂志,2011,14(6):475-477.
[9]Pelletier L,Rebouissou S,Vignijevic D,et al.HNF1αinhibition triggersepithelial-mesenchymal transition in hum an liver cancer cell lines[J].BMC Cancer,2011,10(11):427.
[10]Zhao X L,Sun T,Che N,et al.Promotion of hepatocellular carcinomametastasis through matrix metalloproteinase activation by epithelial-mesenchymal transition regulator Twist1[J].J Cell Mol Med,2011,15(3):691-700.
[11]Whittaker S,Marais R,Zhu A X.The role of signaling pathways in the development and treatment of hepatocellular carcinoma[J].Oncogene,2010,29(36):4989-5005.
[12]顾红兵,李继坤.Hedgehog信号通路与肿瘤的关系研究进展[J].Modern Oncology,2011,19(4):808-811.
[13]Ansieau S,Bastid J,Doreau A,et al.Induction of EMT by twist proteins as a collateral effect of tumorpromoting ina-ctivation of premature senescence[J].Cancer Cell,2008,14(1):79-89.
[14]刘敏,熊成名,王子豫,等.丹参酮ⅡA对肝癌细胞MHCC97-H Smad3/Smad7蛋白及上皮间质转化的影响[J].中医药导报,2017,23(10):30-34.
[15]肖跃红.原发性肝癌的中医病机及防治探讨[J].中国中医基础医学杂志,2013,19(6):617-648.
[16]秦英刚,花宝金.花宝金治疗肝癌经验[J].北京中医药,2013,32(1):33-34.
[17]张存义,钱心兰.扶正祛邪法治疗肝癌之体会[J].上海中医药杂志,2000(11):22-23.
[18]刘俊保,刘延庆.原发性肝癌的中医药治疗研究述评[J].中医学报,2013,28(1):11-13.
[19]李川,吕文良,孙桂芝.孙桂芝教授益气活血解毒散结法治疗肝癌学术思想[J].长春中医药大学学报,2012,28(6):1002-1003.
[20]郝丽君.浅谈陈光伟教授扶正抗癌法治疗肿瘤经验[J].中国民族民间医药,2013,22(11):127.
[21]杨柳青,陈光伟,陈建婷.扶正抗癌汤对肝癌小鼠突变型p53基因和巨噬细胞CD_(68)表达的影响[J].陕西中医,2011,32(10):1426-1427.
[22]方瑜,杨柳青,陈光伟.扶正抗癌方对小鼠肝癌细胞转移影响的实验研究[J].陕西中医,2013,34(9):1253-1255.
[23]方瑜,杨柳青,陈光伟.扶正抗癌方对H_(22)移植瘤小鼠凋亡和端粒酶表达影响的实验研究[J].陕西中医,2013,34(10):1423-1425.
[24]方瑜,杨柳青,王松岩,等.扶正抗癌方对H_(22)细胞STAT3通路的影响[J].陕西中医,2014,35(5):618-619.
[25]方瑜,张淑珍,陈光伟.扶正抗癌方抑制HepG2细胞诱导EMT机理探讨[J].陕西中医药大学学报,2016,39(6):121-124.
[26]彭凯丽,陈宏辉.上皮间质细胞转化在肿瘤侵袭转移及化疗耐药中的作用[J].实用医药杂志,2018,35(1):85-87.
[27]迟笑怡,胡凯文,周天.盐酸川芎嗪对小鼠Lewis肺癌细胞转移及上皮-间质转化的影响[J].中医药导报,2018,24(11):9-15,20.
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