PMID- 26362062 OWN - NLM STAT- MEDLINE DCOM- 20160523 LR - 20231213 IS - 1756-9966 (Electronic) IS - 0392-9078 (Print) IS - 0392-9078 (Linking) VI - 34 IP - 1 DP - 2015 Sep 11 TI - Activation of SAPK/JNK mediated the inhibition and reciprocal interaction of DNA methyltransferase 1 and EZH2 by ursolic acid in human lung cancer cells. PG - 99 LID - 10.1186/s13046-015-0215-9 [doi] LID - 99 AB - BACKGROUND: Ursolic acid (UA), a pentacyclic triterpenoid, is known to have anti-tumor activity in various cancers including human non small cell lung cancer (NSCLC). However, the molecular mechanisms underlying the action of UA remain largely unknown. METHODS: Cell viability was measured by MTT assays. Apoptosis was analyzed with Annexin V-FITC/PI Apoptosis Detection Kit by Flow cytometry. Western blot analysis was performed to measure the phosphorylation and protein expression of stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK), DNMT1 [DNA (cytosine-5)-methyltransferase 1], enhancer of zeste 2 polycomb repressive complex 2 subunit (EZH2) and SP1. Exogenous expression of SP1 and DNMT1 was carried out by transient transfection assays. RESULTS: We showed that UA inhibited the growth and induced apoptosis of NSCLC cells in the dose- and time-dependent fashion. Furthermore, we found that UA induced phosphorylation of SAPK/JNK and suppressed the protein expression of DNMT1 and EZH2. The inhibitor of SAPK/JNK (SP600125) blocked the UA-reduced expression of DNMT1 and EZH2. In addition, UA suppressed the expression of SP1 protein. Conversely, overexpression of SP1 reversed the effect of UA on DNMT1 and EZH2 expression, and feedback attenuated UA-induced phosphorylation of SAPK/JNK. Moreover, exogenous expression of DNMT1 antagonized the effect of UA on SAPK/JNK signaling, EZH2 protein expression, and NSCLC cell growth. CONCLUSION: Our results show that UA inhibits growth of NSCLC cells through SAPK/JNK-mediated inhibition of SP1; this in turn results in inhibition the expression of DNMT1 and EZH2. Overexpression of DNMT1 diminishes UA-reduced EZH2 protein expression. The negative feedback regulation of SAPK/JNK signaling by SP1 and DNMT1, and the reciprocal interaction of EZH2 and DNMT1 contribute to the overall effects of UA. This study leads to important new insights into the mechanisms by which UA controls growth of NSCLC cells. FAU - Wu, Jingjing AU - Wu J AD - Laboratory of Tumor Molecular Biology and Targeted Therapies, Guangdong Provincial Hospital of Chinese Medicine, The Second Clinical Medical Collage, University of Guangzhou Traditional Chinese Medicine, Guangzhou, 510120, Guangdong Province, P. R. China. FAU - Zhao, Shunyu AU - Zhao S AD - Laboratory of Tumor Molecular Biology and Targeted Therapies, Guangdong Provincial Hospital of Chinese Medicine, The Second Clinical Medical Collage, University of Guangzhou Traditional Chinese Medicine, Guangzhou, 510120, Guangdong Province, P. R. China. FAU - Tang, Qing AU - Tang Q AD - Laboratory of Tumor Molecular Biology and Targeted Therapies, Guangdong Provincial Hospital of Chinese Medicine, The Second Clinical Medical Collage, University of Guangzhou Traditional Chinese Medicine, Guangzhou, 510120, Guangdong Province, P. R. China. FAU - Zheng, Fang AU - Zheng F AD - Laboratory of Tumor Molecular Biology and Targeted Therapies, Guangdong Provincial Hospital of Chinese Medicine, The Second Clinical Medical Collage, University of Guangzhou Traditional Chinese Medicine, Guangzhou, 510120, Guangdong Province, P. R. China. FAU - Chen, YuQin AU - Chen Y AD - Laboratory of Tumor Molecular Biology and Targeted Therapies, Guangdong Provincial Hospital of Chinese Medicine, The Second Clinical Medical Collage, University of Guangzhou Traditional Chinese Medicine, Guangzhou, 510120, Guangdong Province, P. R. China. FAU - Yang, LiJun AU - Yang L AD - Laboratory of Tumor Molecular Biology and Targeted Therapies, Guangdong Provincial Hospital of Chinese Medicine, The Second Clinical Medical Collage, University of Guangzhou Traditional Chinese Medicine, Guangzhou, 510120, Guangdong Province, P. R. China. FAU - Yang, Xiaobing AU - Yang X AD - Department of Medical Oncology, Guangdong Provincial Hospital of Chinese Medicine, The Second Clinical Medical Collage, University of Guangzhou Traditional Chinese Medicine, Guangzhou, Guangdong Province, 510120, P. R. China. FAU - Li, Liuning AU - Li L AD - Department of Medical Oncology, Guangdong Provincial Hospital of Chinese Medicine, The Second Clinical Medical Collage, University of Guangzhou Traditional Chinese Medicine, Guangzhou, Guangdong Province, 510120, P. R. China. FAU - Wu, WanYin AU - Wu W AD - Department of Medical Oncology, Guangdong Provincial Hospital of Chinese Medicine, The Second Clinical Medical Collage, University of Guangzhou Traditional Chinese Medicine, Guangzhou, Guangdong Province, 510120, P. R. China. FAU - Hann, Swei Sunny AU - Hann SS AD - Laboratory of Tumor Molecular Biology and Targeted Therapies, Guangdong Provincial Hospital of Chinese Medicine, The Second Clinical Medical Collage, University of Guangzhou Traditional Chinese Medicine, Guangzhou, 510120, Guangdong Province, P. R. China. swhan2010@live.com. AD - , No. 55, Neihuan West Road, Higher Education Mega Center, Panyu District, Guangzhou, Guangdong Province, 510006, P. R. China. swhan2010@live.com. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20150911 PL - England TA - J Exp Clin Cancer Res JT - Journal of experimental & clinical cancer research : CR JID - 8308647 RN - 0 (Antineoplastic Agents) RN - 0 (Triterpenes) RN - EC 2.1.1.37 (DNA (Cytosine-5-)-Methyltransferase 1) RN - EC 2.1.1.37 (DNA (Cytosine-5-)-Methyltransferases) RN - EC 2.1.1.37 (DNMT1 protein, human) RN - EC 2.1.1.43 (EZH2 protein, human) RN - EC 2.1.1.43 (Enhancer of Zeste Homolog 2 Protein) RN - EC 2.1.1.43 (Polycomb Repressive Complex 2) RN - EC 2.7.11.24 (Mitogen-Activated Protein Kinases) SB - IM MH - Antineoplastic Agents/*pharmacology MH - Carcinoma, Non-Small-Cell Lung/*drug therapy MH - Cell Line, Tumor MH - DNA (Cytosine-5-)-Methyltransferase 1 MH - DNA (Cytosine-5-)-Methyltransferases/*metabolism MH - Drug Screening Assays, Antitumor MH - Enhancer of Zeste Homolog 2 Protein MH - Humans MH - Lung Neoplasms/*drug therapy MH - Mitogen-Activated Protein Kinases/metabolism MH - Phosphorylation MH - Polycomb Repressive Complex 2/*metabolism MH - Protein Processing, Post-Translational MH - Signal Transduction MH - Triterpenes/*pharmacology MH - Ursolic Acid PMC - PMC4567809 EDAT- 2015/09/13 06:00 MHDA- 2016/05/24 06:00 PMCR- 2015/09/11 CRDT- 2015/09/13 06:00 PHST- 2015/07/21 00:00 [received] PHST- 2015/09/01 00:00 [accepted] PHST- 2015/09/13 06:00 [entrez] PHST- 2015/09/13 06:00 [pubmed] PHST- 2016/05/24 06:00 [medline] PHST- 2015/09/11 00:00 [pmc-release] AID - 10.1186/s13046-015-0215-9 [pii] AID - 215 [pii] AID - 10.1186/s13046-015-0215-9 [doi] PST - epublish SO - J Exp Clin Cancer Res. 2015 Sep 11;34(1):99. doi: 10.1186/s13046-015-0215-9.