| 摘要: |
| 目的：观察过氧化物酶体增长因子活化受体(peroxisome proliferator-activated receptor，PPAR)γ 激动剂——曲格列酮体外对肝癌细胞生长及β-catenin信号通路的影响，初步探讨其可能的抗肝癌机制。方法：体外培养肝癌细胞HepG2，以MTT法测定不同浓度曲格列酮（5、10、20、40、80、100 μmol/L）作用120 h后细胞存活率，并与正常培养细胞（对照组）作比较；应用流式细胞仪分析曲格列酮（10 μmol/L）处理组及对照组HepG2细胞周期；免疫细胞化学法观察曲格列酮（10 μmol/L）处理组及对照组HepG2细胞β-catenin的亚细胞定位；Western印迹法检测各自cyclin D1和c-myc蛋白的表达。结果：5、10、20、40、80、100 μmol/L曲格列酮作用120 h后，HepG2细胞存活率分别为（96.8±1.2）%、（53.4±1.2）%、（42.3±1.2）%、（31.4±1.0）%、（13.6±0.8）% 和（9.6±0.7）%，组间有统计学差异（P<0.05）。与对照组相比，10 μmol/L曲格列酮处理组细胞G0/G1期细胞比例增加\[（67.6±0.5）% vs （56.3±1.5）%，P＜0.01\]，而S期细胞比例减少\[（20.6±0.5）% vs （25±1.0）%，P＜0.01）\]。对照组细胞β-catenin亚细胞定位于细胞核，而曲格列酮处理组定位于细胞质；Western印迹结果表明曲格列酮处理组细胞c-myc和cyclin D1蛋白表达低于对照组。结论：曲格列酮体外能抑制肝癌细胞生长，且随着浓度的增加，其抑制作用逐渐增强；其抑制作用可能与调控β-catenin信号通路，调节相关靶蛋白表达有关。 |
| 关键词: 过氧化物酶体增长因子活化受体 曲格列酮 肝肿瘤 β-catenin |
| DOI：10.3724/SP.J.1008.2008.00011 |
| 投稿时间：2007-06-18 |
| 基金项目: |
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| Influence of troglitazone on proliferation of human liver cancer cell line HepG2 in vitro and on β-catenin signaling pathway |
| ZHOU Yan-ming1,2,WEN Ying-hao2,KANG Xiao-yan2,QIAN Hai-hua2,LI Dian-qi2,YANG Jia-mei2,LI Bin1,YIN Zheng-feng2* |
| (1.Department of Hepato-Biliary-Pancreato-Vascular Surgery, The First Hospital of Xiamen, Fujian Medical University, Xiamen 361003,China ;2.Laboratory of molecular oncology research, Eastern Hepatobiliary Hospital, Second Military Medical University, Shanghai 200438) |
| Abstract: |
| Objective：To investigate the influence of troglitazone, a potent peroxisome proliferator-activated receptor (PPAR) gamma agonist, on proliferation and β-catenin signaling pathway of human liver cancer cell line HepG2 in vitro, and to discuss its possible anti-cancer mechanism. Methods: HepG2 cells were cultured in vitro and the cell growth was assessed by MTT assay after exposure to different concentrations of troglitazone (5, 10, 20, 40, 80 and 100 μmol/L) for 120 h, and the results were compared with that of the control cells (cultured normally). Flow cytometry was used to assess cell cycle of HepG2 cells treated with troglitazone at 10 μmol/L and of normal control cells. The subcellular location of β-catenin was investigated by immunocytochemistry in troglitazone（10 μmol/L）-treated and control cells. Expression of cyclin D1 and c-myc proteins was examined by Western blotting assay. Results: MTT assay demonstrated that, after treatment with 5, 10, 20, 40, 80 and 100 μmol/L of troglitazone, the cell survival rates were (96.8±1.2）%,（53.4±1.2）%,（42.3±1.2）%, （31.4±1.0）%,（13.6±0.8）% and （9.6±0.7）%, respectively. Compared with control cells, cells treated with 10 μmol/L troglitazone showed an increased proportion of cells at the G0/G1 phase（\[67.6±0.5\]% vs \[56.3±1.5\]%，P＜0.01) and decreased proportion of cells at the S phase（\[20.6±0.5\]% vs \[25±1.0\]%，P＜0.01）. β-catenin was located in the nucleus of the control cells and in the cytoplasm of the troglitazone-treated cells. Western blotting analysis showed that the expression of c-myc and cyclin D1 proteins in troglitazone-treated cells was lower than that in the control cells. Conclusion: Troglitazone can inhibit the proliferation of HepG2 cells in a dose-dependent manner, which may be associated with regulation of the β-catenin signaling pathway and inhibition of target protein expression. |
| Key words: peroxisome proliferator-activated receptor troglitazone liver neoplasms β-catenin |
