PMID- 26850724 OWN - NLM STAT- MEDLINE DCOM- 20161213 LR - 20181211 IS - 1872-7573 (Electronic) IS - 0378-8741 (Linking) VI - 181 DP - 2016 Apr 2 TI - Chinese herbal medicine Xiaoji decoction inhibited growth of lung cancer cells through AMPKalpha-mediated inhibition of Sp1 and DNA methyltransferase 1. PG - 172-81 LID - S0378-8741(16)30041-1 [pii] LID - 10.1016/j.jep.2016.01.041 [doi] AB - ETHNOPHARMACOLOGICAL RELEVANCE: Xiaoji decoction (XJD), which was considered as a Chinese herbal prescription, has been used for cancer treatment, especially lung cancer, for decades to improve quality of life and prolong the patient survival. However, the molecular mechanisms underlying the therapeutic potential have not been well elucidated. MATERIALS AND METHODS: The cell viability was examined by MTT assays. The phosphorylation and expression of AMP-activated protein kinase alpha (AMPKalpha), DNA methyltransferase 1 (DNMT1) and transcription factor Sp1 proteins were assessed by Western Blot. Exogenous expression of Sp1 and DNMT1 were performed by transient transfection methods. The effects of XJD on the growth of xenograft tumors were evaluated by in vivo bioluminescence imaging. RESULTS: We showed that XJD inhibited growth of human non small cell lung cancer (NSCLC) cells in vitro. We also found that XJD increased phosphorylation of AMPKalpha and inhibited protein expression of DNTM1, the latter was not observed in the presence of the inhibitor of AMPK (compound C). Overexpression of DNTM1 reversed the effect of XJD on cell growth. In addition, XJD decreased Sp1 protein expression, which was eliminated by compound C. Conversely, exogenous expressed Sp1 abrogated XJD-inhibited DNTM1 protein expression. Interestingly, exogenous expression of DNMT1 feedback antagonized the XJD-induced phosphorylation of AMPKalpha. In in vivo studies, we found that XJD inhibited tumor growth in xenograft nude mice model, which was accompanied by induction of phosphorylation of AMPKalpha and suppression of DNMT1 protein from xenograft tumors. CONCLUSION: Our results show that XJD inhibits NSCLC cell growth via AMPKalpha-mediated inhibition of transcription of Sp1, followed by the reduction of DNMT1 expression both in vitro and in vivo. The negative feedback regulation loop of AMPKalpha further demonstrates the critical role of DNMT1 in mediating the overall effects of XJD in this process. This study unveils novel molecular mechanism by which XJD controls NSCLC cell growth. CI - Copyright (c) 2016 Elsevier Ireland Ltd. All rights reserved. FAU - Zhao, ShunYu AU - Zhao S AD - Laboratory of Tumor Biology, Guangdong Provincial Hospital of Chinese Medicine, The Second Clinical Medical Collage, University of Guangzhou Traditional Chinese Medicine, Guangzhou, Guangdong Province 510120, China. FAU - Wu, Jingjing AU - Wu J AD - Laboratory of Tumor Biology, Guangdong Provincial Hospital of Chinese Medicine, The Second Clinical Medical Collage, University of Guangzhou Traditional Chinese Medicine, Guangzhou, Guangdong Province 510120, China. FAU - Tang, Qing AU - Tang Q AD - Laboratory of Tumor Biology, Guangdong Provincial Hospital of Chinese Medicine, The Second Clinical Medical Collage, University of Guangzhou Traditional Chinese Medicine, Guangzhou, Guangdong Province 510120, China. FAU - Zheng, Fang AU - Zheng F AD - Laboratory of Tumor Biology, Guangdong Provincial Hospital of Chinese Medicine, The Second Clinical Medical Collage, University of Guangzhou Traditional Chinese Medicine, Guangzhou, Guangdong Province 510120, China. FAU - Yang, LiJun AU - Yang L AD - Laboratory of Tumor Biology, Guangdong Provincial Hospital of Chinese Medicine, The Second Clinical Medical Collage, University of Guangzhou Traditional Chinese Medicine, Guangzhou, Guangdong Province 510120, China. FAU - Chen, YuQin AU - Chen Y AD - Laboratory of Tumor Biology, Guangdong Provincial Hospital of Chinese Medicine, The Second Clinical Medical Collage, University of Guangzhou Traditional Chinese Medicine, Guangzhou, Guangdong Province 510120, 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, China. FAU - Hann, Swei Sunny AU - Hann SS AD - Laboratory of Tumor Biology, Guangdong Provincial Hospital of Chinese Medicine, The Second Clinical Medical Collage, University of Guangzhou Traditional Chinese Medicine, Guangzhou, Guangdong Province 510120, China. Electronic address: 18718844345@163.com. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20160202 PL - Ireland TA - J Ethnopharmacol JT - Journal of ethnopharmacology JID - 7903310 RN - 0 (Drugs, Chinese Herbal) RN - 0 (Sp1 Transcription Factor) RN - 8XXD6IS03Y (xiaoji) RN - 9007-49-2 (DNA) RN - EC 2.1.1.- (Methyltransferases) RN - EC 2.7.11.31 (AMP-Activated Protein Kinases) SB - IM MH - A549 Cells MH - AMP-Activated Protein Kinases/*metabolism MH - Animals MH - Apoptosis/drug effects MH - Carcinoma, Non-Small-Cell Lung/drug therapy/metabolism MH - Cell Line, Tumor MH - Cell Proliferation/drug effects MH - Cell Survival/drug effects MH - DNA/*metabolism MH - Drugs, Chinese Herbal/*pharmacology MH - Female MH - Gene Expression Regulation, Neoplastic/drug effects MH - Humans MH - Lung Neoplasms/*drug therapy/metabolism MH - Methyltransferases/*metabolism MH - Mice MH - Mice, Nude MH - Phosphorylation/drug effects MH - Signal Transduction/drug effects MH - Sp1 Transcription Factor/*metabolism OTO - NOTNLM OT - AMPKalpha OT - DNMT1 OT - NSCLC OT - Sp1 OT - Xiaoji decoction EDAT- 2016/02/07 06:00 MHDA- 2016/12/15 06:00 CRDT- 2016/02/07 06:00 PHST- 2015/06/19 00:00 [received] PHST- 2016/01/29 00:00 [revised] PHST- 2016/01/30 00:00 [accepted] PHST- 2016/02/07 06:00 [entrez] PHST- 2016/02/07 06:00 [pubmed] PHST- 2016/12/15 06:00 [medline] AID - S0378-8741(16)30041-1 [pii] AID - 10.1016/j.jep.2016.01.041 [doi] PST - ppublish SO - J Ethnopharmacol. 2016 Apr 2;181:172-81. doi: 10.1016/j.jep.2016.01.041. Epub 2016 Feb 2.