MEDLINE/PubMed Journal Browser Search

Pubmed for Handhelds

PUBMED FOR HANDHELDS

Search MEDLINE/PubMed

Title: [Transforming growth factor beta regulates hepatic progenitor cells migration via PI3K/AKT/mTOR/p70S6K pathway].
Author: Pu XH, Li F, Miao XL, Ye JL, Lu LG.
Journal: Zhonghua Gan Zang Bing Za Zhi; 2018 Sep 20; 26(9):680-685. PubMed ID: 30481866.
Abstract:
Objective: To investigate the effect and mechanism of transforming growth factor β (TGFβ) on the migration ability of hepatic progenitor cells in vitro. Methods: Primary hepatic progenitor cells of male wild-type C57BL/6J mice were isolated by two-step perfusion method and stimulated with different concentrations of TGFβ .The morphological changes were observed under phase -contrast microscopy. The effects of TGFβ on migration ability of hepatic progenitor cells were evaluated by scratch test and transwell method. Expression profiling and signaling phospho antibody array detected the signaling pathways involved in the regulation of TGFβ on hepatic progenitor cells. Protein level of PI3K/AKT/mTOR/p70S6K signaling pathway and the localization of each signaling molecules in hepatic progenitor cells were detected. Data comparison between the two groups was performed by independent sample t-test. One-way ANOVA was used for data comparison between multiple groups. Results: TGFβ made the liver progenitor cells from oval to long spindle type. Scratch test showed that the scratch healing rates of 24 h control group, and 2 ng/ml and 10 ng/ml TGF-beta groups were 36.48% ± 4.37%, 57.35% ± 4.60%, and 73.14% ± 5.02% (F = 65.87, P < 0.01), respectively. Transwell test showed that the number of migrating cells in 24 h control group, 2 ng/ml and 10 ng/ml TGF-beta groups were 127 ± 16, 230 ± 18, and 385 ±36 (F = 94.99, P < 0.01), respectively. The results of expression profiling showed that TGFβ regulates gene expression in hepatic progenitor cells, and differentially expressed genes participate in the PI3K-AKT signaling pathway. Signaling phospho antibody array and western blot showed that TGFβ regulated PI3K/AKT/mTOR/p70S6K signaling pathway in hepatic progenitor cells. Concurrently, immunofluorescence assay showed phosphorylation (p) 70s6k, p AKT1 and PI3K and F-actin co-localizations. Conclusion: TGFβ can promote hepatic progenitor cell migration through PI3K/AKT/mTOR/p70S6K pathway, and p70S6K, pAKT1 and PI3K signaling molecules are involved in the regulation of morphology and migration of liver progenitor cells. 目的： 探讨转化生长因子（TGF）β在体外对肝祖细胞迁移能力的影响及其作用机制。 方法： 两步灌注法分离雄性野生型C57BL/6J小鼠原代肝祖细胞，用不同浓度TGFβ刺激，相差显微镜下观察细胞形态；划痕实验和Transwell实验评估TGFβ对肝祖细胞迁移能力的影响；表达谱测序和信号通路磷酸化抗体检测TGFβ对肝祖细胞调控涉及的信号通路，并检测PI3K/AKT/mTOR/p70S6K信号通路蛋白水平及各信号分子在肝祖细胞中的定位。两组间数据比较用独立样本t检验，多组间数据比较用单因素方差分析。 结果： TGFβ使肝祖细胞由卵圆形变为长梭型，划痕实验显示24 h对照组和2 ng/ml TGFβ、10 ng/ml TGFβ组的划痕愈合率分别为36.48%±4.37%、57.35%±4.60%、73.14%±5.02%（F = 65.87，P < 0.01）。Transwell实验显示24 h对照组和2 ng/ml TGFβ、10 ng/ml TGFβ组的迁移细胞数分别为（127±16）个、（230±18）个、（385±36）个（F = 94.99，P < 0.01）。表达谱测序结果表明TGFβ调控肝祖细胞基因表达，其差异表达基因参与PI3K/AKT信号通路。信号通路磷酸化抗体芯片和Western blot检测结果显示TGFβ能调控肝祖细胞PI3K/AKT/mTOR/p70S6K信号通路。同时免疫荧光实验结果显示磷酸化（p）70S6K、pAKT1和PI3K与F-肌动蛋白共定位。 结论： TGFβ能通过PI3K/AKT/mTOR/p70S6K通路促进肝祖细胞迁移，p70S6K、pAKT1和PI3K信号分子参与调控肝脏祖细胞的形态改变和迁移。.

[Abstract] [Full Text] [Related] [New Search]