摘要
目的:探讨健脾化瘀方对转化生长因子β1(TGF-β1)诱导的肝癌细胞MHCC-97H上皮间质转化(EMT)及侵袭转移能力的影响。方法:将不同浓度(0.5、1、2、4、8μmol/L)的健脾化瘀方作用于肝癌细胞MHCC-97H,采用四甲基偶氮唑盐(MTT)法评估细胞活力;TGF-β1诱导肝癌细胞建立EMT模型,倒置显微镜观察细胞形态的变化;根据加入不同浓度健脾化瘀方干预,将实验分为空白组、EMT组、EMT+健脾化瘀方2 mg/mL组(低浓度组)、EMT+健脾化瘀方4 mg/mL组(高浓度组);划痕法和Transwell实验检测肝癌细胞侵袭/迁移能力;Western Blot法检测EMT标志蛋白E-cadherin、N-cadherin和Vimentin蛋白表达水平。结果:健脾化瘀方能抑制肝癌细胞MHCC-97H的增殖,并呈一定的时间-剂量依赖关系(P<0.01)。显微镜下观察经过TGF-β1诱导后的肝癌细胞形态松散,呈梭形样改变。与空白组比较,EMT组的迁移/侵袭能力显著增强(P<0.01),给予药物干预后,高、低浓度组的迁移/侵袭能力较EMT组显著减弱(P<0.01)。与空白组比较,EMT组细胞中的E-cadherin蛋白表达显著降低(P<0.01),N-cadherin和Vimentin蛋白表达显著升高(P<0.01);高、低浓度组E-cadherin蛋白表达显著上调(P<0.01),而N-cadherin和Vimentin蛋白表达显著下调(P<0.01)。结论:健脾化瘀方能抑制TGF-β1诱导的肝癌细胞MHCC-97H的迁移/侵袭能力,其机制可能与抑制上皮间质转化有关。
Objective:To investigate the effects of Jianpi Huayu Formula on the invasion and epithelial-mesenchymal transition in liver cancer cell line MHCC-97H induced by transforming growth factor-β1.Methods:We used different concentrations(0.5,1,2,4,8 mg/mL)of Jianpi Huayu Formula to intervene hepatocellular carcinoma cell line MHCC-97H and the cell viability was measured by MTT method.Hepatoma cells were induced by TGF-β1 to establish EMT model,and the morphological changes of cells were observed under inverted microscope.According to the intervention of Jianpi Huayu Formula with different concentrations,the experiments were divided into blank control group,EMT group,EMT+2 mg/mL group(low dose group),and EMT+4 mg/mL group(high dose group).Wound healing assays and Transwell test were used to detect the migration and invasion ability of MHCC-97H cell.The expression levels of EMT marker proteins E-cadherin,N-cadherin and Vimentin were detected by Western Blot.Results:Jianpi Huayu Formula could inhibit the proliferation of liver cancer MHCC-97H cells in a time-dose-dependence relationship(P<0.01).The morphology of the cells was observed under a microscope,and the hepatoma cells induced by TGF-β1 were loose in shape and showed spindle-like changes.The scratch test and Transwell test results showed that compared with the blank group,the migration/invasion ability of the EMT group was significantly enhanced(P<0.01).After drug intervention,the migration/invasion ability of low and high dose group was significantly weakened compared with EMT group(P<0.01).Compared with the blank group,the expression of E-cadherin protein in cells in EMT group was significantly decreased(P<0.01),and the protein expressions of N-cadherin and Vimentin were significantly increased(P<0.01).E-cadherin protein expression was significantly up-regulated in low and high dose group(P<0.01),while N-cadherin and Vimentin protein expressions were significantly down-regulated(P<0.01).Conclusion:Jianpi Huayu Formula can inhibit TGF-β1-induced migration/inva
作者
谢春凤
陈清莲
冯坤良
方崇锴
王冀南
钟崇
XIE Chun-fengu;CHEN Qing-lian;FENG Kun-liang;FANG Chong-kai;WANG Ji-nan;ZHONG Chong(First Clinical Medical School,Guangzhou University of Chinese Medicine,Guangzhou 510405,China;Department of General Surgery,Liuzhou Traditional Chinese Medicine Hospital,Liuzhou 545001,China;Lingnan Medical Research Center,Guangzhou University of Chinese Medicine,Guangzhou 510405,China;Department of Hepatobiliary Surgery,The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405,China)
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2022年第4期2270-2274,共5页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国家自然科学基金项目(No.81873303)
广东省自然科学基金面上项目(No.2019A1515011013)
广东省教育厅普通高校重点项目(No.2019KZDXM045)。
关键词
健脾化瘀方
肝细胞癌
上皮间质转化
转化生长因子Β1
Jianpi Huayu Formula
Hepatocellular carcinoma
Epithelial-mesenchymal transition
Transforming growth factor-β1
