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| 1 | Role of autophagy in differential sensitivity of hepatocarcinoma cells to sorafenib显示文摘AIM: To investigate the role of sorafenib(SFN) in autophagy of hepatocellular carcinoma(HCC). We evaluated how SFN affects autophagy signaling pathway in human HCC cell lines. METHODS: Two different human HCC cell lines, Hep3 B and Huh7, were subjected to different concentrations of SFN. Cell viability and onset of apoptosis were determined with colorimetric assay and immunoblotting analysis, respectively. The changes in autophagy-related proteins, including LC3, ULK1, AMPK, and LKB, were determined with immunoblotting analysis in the presence or absence of SFN. To assess autophagic dynamics, autophagic flux was measured with chloroquine, a lysosomal inhibitor. The autophagic responsiveness between different HCC cell lines was compared under the autophagy enhancing conditions.RESULTS: Hep3 B cells were significantly more resistant to SFN than Huh7 cells. Immunoblotting analysis revealed a marked increase in SFN-mediated autophagy flux in Huh7 cells, which was, however, absent in Hep3 B cells. While both starvation and rapamycin enhanced autophagy in Huh7 cells, only rapamycin increased autophagy in Hep3 B cells. Immunoblotting analysis of autophagy initiation proteins showed that SFN substantially increased phosphorylation of AMPK and consequently autophagy in Huh7, but not in Hep3 B cells.CONCLUSION: The autophagic responsiveness to SFN is distinct between Hep3 B and Huh7 cells. Resistance of Hep3 B cells to SFN may be associated with altered autophagy signaling pathways. | Trevan D Fischer Jin-Hee Wang Adrian Vlada Jae-Sung Kim Kevin E Behrns | 2014 | World Journal of Hepatology2014，6，10： | 5 |
| 2 | 新城疫病毒诱导原代鸡胚成纤维细胞自噬的研究显示文摘为掌握新城疫病毒(NDV)在原代鸡胚成纤维细胞(CEF)上诱导细胞自噬的特性,用NDV感染CEF后,以电镜观察自噬体形成、GFP-LC3荧光迁移以及Western-blot检测LC3-Ⅰ/Ⅱ转化判定自噬。p62降解和GFP-LC3向溶酶体的转移判定自噬流。结果显示,在感染后第12~24小时能在细胞中发现大量双层或单层膜结构;转染的GFP-LC3质粒在病毒感染后呈现显著的点状分布,尤其在NDV感染形成的合胞体中明显;且病毒感染后期发生明显的LC3-Ⅰ向LC3-Ⅱ的转化。NDV感染能够诱导p62蛋白的降解,共聚焦结果显示,GFP-LC3在病毒感染中后期发生向溶酶体的转移,说明NDV感染能够诱导完整自噬流的产生。结果表明,NDV感染CEF后能够强烈诱导细胞自噬发生,这为进一步研究新城疫病毒和禽源细胞互作奠定了基础。 | 王晓旭 孙英杰 胡跃 仇旭升 谭磊 宋翠萍 王桂军 丁铲 | 2014 | 中国兽医科学2014，44，5： | 3 |
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