跨膜蛋白173对TGF-β1活化的肝星状细胞增殖与凋亡的影响及机制研究

Effect of TMEM173 on proliferation and apoptosis of hepatic stellate cells activated by TGF-β1 and its mechanisms

目的观察跨膜蛋白173(TMEM173)的表达变化对TGF-β1诱导活化的人肝星状细胞(LX-2)增殖和凋亡的影响及其机制。方法TGF-β1诱导建立LX-2细胞活化模型,分别利用MTT、EdU和FCM技术检测TMEM173的表达变化对活化LX-2细胞增殖和凋亡的影响,并进一步利用WB技术检测其对细胞增殖和凋亡相关通路蛋白表达的影响。结果TGF-β1成功诱导LX-2细胞活化,且与对照组相比,GFP-TMEM173组细胞的增殖率明显升高、且凋亡率明显降低,而TMEM173-siRNA组细胞的结果则相反;另外,与对照组相比,GFP-TMEM173组细胞中PCNA和Bcl-2的蛋白表达水平明显上调,且Bax、cleaved-caspase-3和cleaved-PARP的表达水平明显下调,而TMEM173-siRNA组细胞的结果则相反。结论TMEM173对活化LX-2细胞的增殖和凋亡具有调控功能,即其过表达能够促进LX-2细胞增殖和抑制细胞凋亡,反之则抑制LX-2细胞增殖和促进细胞凋亡,表明TMEM173可能在肝星状细胞增殖与活化过程中扮演着积极的角色,是肝纤维化基因治疗的潜在靶点。

Aim To investigate the effect of TMEM173 on proliferation and apoptosis and its mechanism through overexpressing and silencing of TMEM173 in TGF-β1 induced LX-2 cells respectively.MethodsThe activation of LX-2 cells was induced by TGF-β1,and the effects of TMEM173 on the proliferation and apoptosis of LX-2 cells were detected by MTT,EdU and FCM.Furthermore,the expressions of proliferation and apoptosis-related signaling pathway protein were detected by Western blot.Results TGF-β1 induced the activation of LX-2 cells successfully,and the results of MTT and FCM showed that compared with control groups,the proliferation rate of GFP-TMEM173 group significantly increased,and the apoptotic rate decreased significantly,while the proliferation rate of TMEM173-siRNA group significantly decreased,and the apoptotic rate markedly increased.WB results showed that compared with control group,the protein expression levels of PCNA and Bcl-2 in GFP-TMEM173 group were significantly up-regulated,and the expression levels of Bax,cleaved-Caspase-3 and cleaved-PARP were significantly down-regulated in GFP-TMEM173 group,while the results in TMEM173-siRNA group were the opposite.Conclusions TMEM173 can regulate the proliferation and apoptosis of activated LX-2 cells,and its overexpression can promote cell proliferation and inhibit apoptosis significantly,while its silencing can get the opposite results,indicating that TMEM173 may play an active role in the proliferation and activation of hepatic stellate cells,and it is a potential target for gene therapy of hepatitis and liver fibrosis.