摘要
目的探讨Zeste基因增强子同源物2(enhancer of zeste homolog 2,EZH2)表达沉默对大鼠肝星状细胞HSC-T6细胞增殖活化的影响,及其部分调控机制。方法应用EZH2的抑制剂DZNep(3-Deazaneplanocin A)作用于TGF-β1诱导活化的HSC-T6细胞,采用Western blot法检测蛋白EZH2、p-ERK、p-AKT和α-SMA的表达;根据EZH2的碱基序列设计并合成小干扰RNA(small interfering RNA,si RNA),通过脂质体LipofectamineTM2000转染到HSC-T6细胞内,应用四甲基偶氮唑盐(MTT)法检测HSC-T6细胞的增殖变化,Western blot法检测蛋白EZH2、p-ERK、p-AKT和α-SMA的表达。结果将DZNep加入TGF-β1诱导活化的HSC-T6细胞后,EZH2蛋白水平明显降低,同时p-ERK、pAKT和α-SMA蛋白水平亦明显降低;将EZH2-si RNA转染活化的HSC-T6细胞内,HSC-T6细胞的增殖可明显被抑制,同时EZH2、p-ERK、p-AKT和α-SMA蛋白水平亦明显降低。结论抑制EZH2的表达可明显抑制HSC-T6细胞的增殖活化,EZH2可能是潜在的治疗肝纤维化的靶点。
Aim To investigate the effects of cell pro- liferation and activation in HSC-T6 cells by inhibiting the expression of EZH2, and its partial relevant mech- anism. Methods By introducing the inhibitor DZNep in activated HSC-T6 cells stimulated by TGF-β1, the protein expression levels of EZH2, p-ERK, p-AKT and α-SMA were detected by Western blot. The siRNA targeting EZH2 was designed and synthesized according to its nucleotide sequence, and their corresponding ex- pression vectors were constructed and transfected into HSC-T6 ceils with LipofectamineTM 2000. The prolifer- ation of HSC-T6 cells was determined by MTT. And the protein expression levels of EZH2, p-ERK, p-AKT and α-SMA were measured by Western blot. Results By introducing the inhibitor DZNep in activated HSC-T6 cells stimulated by TGF-β1, it effectively de- creased the protein levels of EZH2 and also the protein levels of p-ERK, p-AKT and α-SMA. By introducing EZH2-siRNA in activated HSC-T6 cells, it effectively inhibited the cell proliferation, and also the protein levels of EZH2, p-ERK, p-AKT and α-SMA. Conclu- sion Silencing EZH2 expression inhibits HSC-T6 cell proliferation and activation, and EZH2 may be a poten- tial therapeutic target gene for hepatic fibrosis.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2015年第8期1061-1065,共5页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No 81273526)
博士点基金资助项目(No 20123420120001)
安徽省教育厅基金资助项目(No KJ2012A156)
安徽省自然科学基金资助项目(No 1308085MH145)
安徽医科大学基金资助项目(No XJ201118)
