软坚护肝片含药血清对肝癌细胞迁移侵袭作用及机制  

Research of Mechanisms of Ruanjian Hugan Table on Inhibition of Hepatocellular Carcinoma Migration and the Epithelial Mesenchymal Cell Transformation

作　　者：孙振 李书[2] 程彬彬 周晓东 梁水庭[4] 陈兴东 SUN Zhen;LI Shu;CHENG Binbin;ZHOU Xiaodong;LIANG Shuiting;CHEN Xingdong(Four Districts for Convalescence of Air Force Hangzhou Special Service Recuperation Center,Nanjing 211131,Jiangsu,China;Baoshan Branch of Shuguang Affiliated Hospital of Shanghai University of Traditional Chinese Medicine,Shanghai 201900,China;Naval Medical University,Shanghai 200433,China;The First Affiliated Hospital of Guangxi Medical University,Nanning 530021,Guangxi,China)

机构地区：[1]空军杭州特勤疗养中心疗养四区,江苏南京211131 [2]上海中医药大学附属曙光医院宝山分院,上海201900 [3]海军军医大学,上海200433 [4]广西医科大学第一附属医院,广西南宁530021

出　　处：《辽宁中医药大学学报》2020年第3期22-25,共4页Journal of Liaoning University of Traditional Chinese Medicine

基　　金：国家自然科学基金(81603458);空军杭州特勤疗养中心科研课题(19TLMS010);南京总医院军事医学课题(YYMS2014013);南京总医院院管课题(2015068,2016083);江苏省药学会-奥赛康医院药学科研项目(201521)。

摘　　要：目的观察软坚护肝片含药血清对肝癌细胞迁移侵袭的抑制作用及其对细胞上皮间质转化(EMT)的影响。方法采用不同剂量软坚护肝片制备软坚护肝片含药血清,处理体外培养的人肝癌Huh7细胞株,噻唑蓝还原法(MTT)检测肝癌细胞的增殖,采用小室迁移穿模(Transwell)实验检测肝癌细胞的迁移、侵袭能力,蛋白质印迹法(Western blot)、荧光定量逆转录-聚合酶链反应(Reverse Transcription-Polymerase Chain Reaction,RTPCR)检测E-cadherm、Vimentin表达。结果软坚护肝片含药血清能显著抑制Huh7细胞增殖,与空白对照组相比差异均有统计学意义(P<0.01)。转化生长因子-β1(TGF-β1)处理后,Huh7细胞迁移数目较对照组显著增加(P<0.01)。而低、高剂量软坚护肝片含药血清组较TGF-β1组显著降低(P<0.01)。TGF-β1处理后,E-cadherin蛋白及mRNA均较对照组显著降低,而软坚护肝片含药血清组E-cadherin蛋白及mRNA表达水平均较TGF-β1组升高。而TGF-β1处理后Vimentin蛋白及mRNA表达水平较对照组显著升高,而软坚护肝片含药血清能显著抑制Vimentin蛋白及mRNA表达。结论软坚护肝片含药血清能够显著抑制肝癌细胞的增殖、迁移和侵袭能力,抑制肝癌细胞EMT是其发挥抗肿瘤作用的机制之一。Objective To investigate the mechanisms of Ruanjian Hugan Table on inhibition of hepatocellular carcinoma(HCC)migration and the epithelial mesenchymal cell transformation(EMT).Methods The RJHG Tablet-medicated serum was prepared with different doses of RJHG Tablet,and the Huh7 cell lines of human liver cancer cultured in vitro were treated.The effect of RJHG Tablet-medicated serum on the growth of Huh7 cells was evaluated by MTT assay in vitro.Transwell assay was used to detect the migration and invasion ability of liver cancer cells.the expression of E-cadherm and Vimentin was determined using Western blot and fluorescence quantitative RT-PCR.Results RJHG Tablet-medicated serum significantly could significantly inhibit the proliferation of Huh7 cells,and the difference was statistically significant compared with the blank control group(P<0.01).After TGF-β1 treatment,the number of Huh7 cell migration was significantly increased compared with the control group(P<0.01),and compared with TGF-β1 group,low and high dose RJHG Tablet-medicated serum significantly reduced in serum group(P<0.01).After TGF-β1 treatment,E-cadherin protein and mRNA levels were significantly lower than those in the control group,while E-cadherin protein and mRNA levels were significantly higher in the RJHG Tablet-medicated serum than in the TGF-β1 group.The expression level of Vimentin protein and mRNA in TGF-β1 treatment was significantly higher than that in the control group,while RJHG Tablet-medicated serum could significantly inhibit the expression of Vimentin protein and mRNA.Conclusion In conclusion,our data suggest that RJHG Tablet-medicated serum can significantly inhibit the proliferation,migration and invasion of liver cancer cells,and the inhibition of liver cancer cell EMT is one of the anti-tumor mechanisms.

关 键 词：软坚护肝片 原发性肝癌 上皮间质转化 转移 

分 类 号：R735.7[医药卫生—肿瘤]