摘要
目的分析FAM172A蛋白对HepG2细胞增殖的影响及相关信号通路,为肝癌治疗新靶点提供理论基础。方法收集空军特色医学中心(原中国人民解放军空军总医院)肝胆外科2016年3月至2018年2月11例患者手术切除的肝组织,包括正常肝组织、感染乙型肝炎病毒(HBV)的肝组织及肝癌组织。HepG2细胞随机分为实验组和对照组。实验组共同转染FAM172A蛋白质粒和荧光蛋白质粒,对照组无任何处理,然后激光共聚焦显微镜观察。Western印迹测定LO2、HepG2、HerpG2.2.15细胞中FAM172A表达。不同浓度FAM172A重组蛋白溶液与HepG2细胞共孵育不同时间,以PBS为对照,流式细胞仪检测细胞周期和细胞计数,Western印迹检测Notch信号和细胞周期蛋白表达。结果FAM172A蛋白位于HepG2细胞的内质网,在正常肝组织明显表达,在感染HBV患者肝组织表达下降。随着FAM172A浓度的增加,对肝癌细胞HepG2增殖的抑制作用越明显,呈浓度依赖性。100.0pg/LFAM172A与HepG2细胞共孵育48h后,在S期的占2.27%,G1期的占77.49%,细胞完全阻滞于G1/S期。电泳结果显示,不同浓度FAM172A干预后,仅Notch3蛋白和细胞周期蛋白E相对表达增加。结论FAM172A可能是一种新的抑癌基因,通过激活Notch3信号通路和上调细胞周期蛋白E表达,抑制肝癌细胞增殖。
Objective To analyze the effect of FAM172A protein on HepG2 cell proliferation and related signaling pathways, and provide a theoretical basis for the new target of liver cancer treatment. Methods The liver tissue was removed from March 2016 to February 2018 in the Air Force Characteristic Medical Center (formerly the People's Liberation Army Air Force General Hospital). The liver tissues were surgically resected, including normal liver tissue and liver infected with hepatitis B virus (HBV) tissue and liver cancer tissue, HepG2 cells were randomly divided into experimental group and control group. The experimental group was co-transfected with FAM172A protein granules and fluorescent protein-labeled plasmids, and the control group was left without any treatment. The position of FAM172A protein in HepG2 cells was observed by laser confocal microscopy. The content of FAM172A in LO2, HepG2 and HerpG2.2.15 cells was determined. Different concentrations of FAM172A recombinant protein solution were co-incubated with HepG2 cells for different time. Cell cycle and cell count were detected by flow cytometry, and Notch signal and cyclin expression were detected by Western blot. Results The FAM172A protein was located in the endoplasmic reticulum of HepG2 cells and was clearly expressed in normal liver tissues, and was decreased in liver tissues of patients with HBV infection. With the increase of the concentration of FAM172A, the inhibitory effect on the proliferation of hepatoma cells was more obvious, and it was concentration-dependent. After HepG2 cells were treated with 100 pg/L FAM172A for 48 hours, the percentage in the S phase decreased to 2.27% and in the G1 phase increased to 77.49%, the cell cycle was completely blocked in the G1/S phase. Electrophoresis results showed that only the relative expression of Notch3 protein and cyclin E increased. Conclusion FAM172A may be a novel tumor suppressor gene that inhibits the proliferation of hepatoma cells by activating Notch3 signaling pathway and up-regulating the expression of cyclin E.
作者
叶军
韩山山
冯志强
Ye Jun;Han shanshan;Feng Zhiqiang(Clinical College of Air Force,Anhui Medical University,Hefei 230032,China;Department of Hepatobiliary Surgery,Air Force Characteristic Medical Center,PLA,Beijing 100142,China;Beijing Chaoyang Emergengcy Medical Center,Beijing 100142,China)
出处
《中华肝胆外科杂志》
CAS
CSCD
北大核心
2019年第6期443-447,共5页
Chinese Journal of Hepatobiliary Surgery