Resveratrol analogue 3,4,4＇-trihydroxy-trans-stilbene nduces apoptosis and autophagy in human non-small-cell lung cancer cells in vitro  

作　　者：Lu ZHANG Fang DAI Pan-long SHENG Zhi-qiang CHEN Qi-ping XU Yu-qi GUO 

机构地区：[1]College of Bioengineering, He-nan University of Technology, Lianhua Street, Zhengzhou 450001, China [2]State Key Laboratory of Applied Organic Chemistry, Lanzhou University, Lanzhou 730000, China [3]Department of Obstetrics and Gynecology, The Third Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China

出　　处：《Acta Pharmacologica Sinica》2015年第10期1256-1265,共10页中国药理学报（英文版）

基　　金：Acknowledgements This study was financially supported by the National Natural Science Foundation of China （No 31101001, 31471296, and 21372109）, the Program for Science and Technology Innova- tion Talents in Universities of He-nan Province （No 15HASTIT031）, the Key Project of He-nan Educational Committee （No 14A180028）, the Foundation for University Key Teacher by He-nan Educational Committee （No 2013GGJS-078） and the Technological innovation incubator program from He-nan University of Technology （No 11CXRC12）.

摘　　要：Aim： To investigate the effects of 3,4,4＇-trihydroxy-trans-stilbene （3,4,4＇-THS）, an analogue of resveratrol, on human non-small-cell lung cancer （NSCLC） cells in vitro.Methods： Cell viability of NSCLC A549 cells was determined by MTT assay. Cell apoptosis was evaluated using flow cytometry and TUNEL assay. Cell necrosis was evaluated with LDH assay. The expression of apoptosis- or autophagy-associated proteins was measured using Western blotting. The formation of acidic compartments was detected using AO staining, neutral red staining and Lysotracker-Red staining. LC3 punctae were analyzed with fluorescence microscopy. Results： Treatment with 3,4,4＇-THS （10-80 μmol/L） concentration-dependently inhibited the cell viability. It did not cause cell necrosis, but induced apoptosis accompanied by up-regulation of cleavaged PARP, caspase3/9 and Bax, and by down-regulation of Bcl-2 and surviving. It also increased the formation of acidic compartments, LC3-II accumulation and GFP-LC3 labeled autophagosomes in the cells. It inhibited the mTOR-dependent pathway, but did not impair autophagic flux. 3,4,4＇-THS-induced cell death was enhanced by the autophagy inhibitors 3-MA （5 mmol/L） or Wortmannin （2 μmol/L）. Moreover, 3,4,4＇-THS treatment elevated the ROS levels in the cells, and co-treatment with 3-MA further elevated the ROS levels. 3,4,4＇-THS-induced apoptosis and autophagy in the cells was attenuated by NAC （10 mmol/L）.Conclusion： 3,4,4＇-THS induces both apoptosis and autophagy in NSCLC A549 cells in vitro. Autophagy inhibitors promote 3,4,4＇-THS-induced apoptosis of A549 cells, thus combination of 3,4,4＇-THS and autophagy inhibitor provides a promising strategy for NSCLC treatment.Aim： To investigate the effects of 3,4,4＇-trihydroxy-trans-stilbene （3,4,4＇-THS）, an analogue of resveratrol, on human non-small-cell lung cancer （NSCLC） cells in vitro.Methods： Cell viability of NSCLC A549 cells was determined by MTT assay. Cell apoptosis was evaluated using flow cytometry and TUNEL assay. Cell necrosis was evaluated with LDH assay. The expression of apoptosis- or autophagy-associated proteins was measured using Western blotting. The formation of acidic compartments was detected using AO staining, neutral red staining and Lysotracker-Red staining. LC3 punctae were analyzed with fluorescence microscopy. Results： Treatment with 3,4,4＇-THS （10-80 μmol/L） concentration-dependently inhibited the cell viability. It did not cause cell necrosis, but induced apoptosis accompanied by up-regulation of cleavaged PARP, caspase3/9 and Bax, and by down-regulation of Bcl-2 and surviving. It also increased the formation of acidic compartments, LC3-II accumulation and GFP-LC3 labeled autophagosomes in the cells. It inhibited the mTOR-dependent pathway, but did not impair autophagic flux. 3,4,4＇-THS-induced cell death was enhanced by the autophagy inhibitors 3-MA （5 mmol/L） or Wortmannin （2 μmol/L）. Moreover, 3,4,4＇-THS treatment elevated the ROS levels in the cells, and co-treatment with 3-MA further elevated the ROS levels. 3,4,4＇-THS-induced apoptosis and autophagy in the cells was attenuated by NAC （10 mmol/L）.Conclusion： 3,4,4＇-THS induces both apoptosis and autophagy in NSCLC A549 cells in vitro. Autophagy inhibitors promote 3,4,4＇-THS-induced apoptosis of A549 cells, thus combination of 3,4,4＇-THS and autophagy inhibitor provides a promising strategy for NSCLC treatment.

关 键 词：3 4 4-trihydroxy-trans-stilbene RESVERATROL non-small-cell lung cancer APOPTOSIS autopha~  mTOR 3-MA WORTMANNIN NAC 

分 类 号：Q255[生物学—细胞生物学] Q782