title: Sann-Joong-Kuey-Jian-Tang induces autophagy in HepG2 cells via regulation of the phosphoinositide-3 kinase/Akt/mammalian target of rapamycin and p38 mitogen-activated protein kinase pathways.
abstract: Sann-Joong-Kuey-Jian-Tang (SJKJT), a traditional Chinese medicine, was previously reported to induce autophagy and inhibit the proliferation of the human HepG2 hepatocellular carcinoma cell line via an extrinsic pathway. In the present study, the effects of SJKJT-induced autophagy and the cytotoxic mechanisms mediating these effects were investigated in HepG2 cells. The cytotoxicity of SJKJT in the HepG2 cells was evaluated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The results demonstrated that the half-maximal inhibitory concentration of SJKJT was 2.91 mg/ml at 24 h, 1.64 mg/ml at 48 h and 1.26 mg/ml at 72 h. The results of confocal fluorescence microscopy indicated that SJKJT resulted in the accumulation of green fluorescent protein-LC3 and vacuolation of the cytoplasm. Flow cytometric analysis revealed the accumulation of acidic vesicular organelles. Furthermore, western blot analysis, used to determine the expression levels of autophagy-associated proteins, demonstrated that the HepG2 cells treated with SJKJT exhibited LC3B-I/LC3B-II conversion, increased expression levels of Beclin, Atg-3 and Atg-5 and reduced expression levels of p62 and decreased signaling of the phosphoinositide-3 kinase/Akt/mammalian target of rapamycin and the p38 mitogen-activated protein kinase pathways. Taken together, these findings may assist in the development of novel chemotherapeutic agents for the treatment of malignant types of liver cancer.
authors.affiliation: Transplant Medicine and Surgery Research Centre, Changhua Christian Hospital, Changhua 50006, Taiwan, R.O.C.
authors.forename: Wan-Ling
authors.initials: WL
authors.lastname: Chuang
authors.affiliation: Department of Surgery, Changhua Christian Hospital, Changhua 50006, Taiwan, R.O.C.
authors.forename: Chin-Cheng
authors.initials: CC
authors.lastname: Su
authors.affiliation: Transplant Medicine and Surgery Research Centre, Changhua Christian Hospital, Changhua 50006, Taiwan, R.O.C.
authors.forename: Ping-Yi
authors.initials: PY
authors.lastname: Lin
authors.affiliation: Institute of Biomedical Science, National Chung-Hsing University, Taichung 40227, Taiwan, R.O.C.
authors.forename: Chi-Chen
authors.initials: CC
authors.lastname: Lin
authors.affiliation: Transplant Medicine and Surgery Research Centre, Changhua Christian Hospital, Changhua 50006, Taiwan, R.O.C.
authors.forename: Yao-Li
authors.initials: YL
authors.lastname: Chen
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism; D001343:Autophagy / Q000187:drug effects*; D000071187:Autophagy-Related Protein 5; D000071183:Autophagy-Related Proteins; D000071186:Beclin-1; D002470:Cell Survival / Q000187:drug effects; D015536:Down-Regulation / Q000187:drug effects; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D056945:Hep G2 Cells; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism; D018613:Microscopy, Confocal; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D058570:TOR Serine-Threonine Kinases / Q000378:metabolism; D044763:Ubiquitin-Conjugating Enzymes / Q000378:metabolism; D015854:Up-Regulation / Q000187:drug effects; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism*
chemical_list: D000970:Antineoplastic Agents; D051017:Apoptosis Regulatory Proteins; D000071187:Autophagy-Related Protein 5; D000071183:Autophagy-Related Proteins; D000071186:Beclin-1; D004365:Drugs, Chinese Herbal; D008565:Membrane Proteins; D008869:Microtubule-Associated Proteins; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt; D058570:TOR Serine-Threonine Kinases; D044763:Ubiquitin-Conjugating Enzymes; D048051:p38 Mitogen-Activated Protein Kinases; C502778:ATG5 protein, human; C491997:BECN1 protein, human; C477012:MAP1LC3A protein, human; C584126:sann-joong-kuey-jian-tang; C455621:ATG3 protein, human
references.citation: Biochim Biophys Acta. 2011 Sep;1813(9):1619-33
references.pmid: 21167873
references.citation: Cell Biochem Funct. 2013 Jul;31(5):427-33
references.pmid: 23086777
references.citation: Mol Med Rep. 2009 Sep-Oct;2(5):707-11
references.pmid: 21475889
references.citation: Cancer Res. 2001 Jan 15;61(2):439-44
references.pmid: 11212227
references.citation: Autophagy. 2007 Nov-Dec;3(6):635-7
references.pmid: 17786026
references.citation: Drug Resist Updat. 2001 Aug;4(4):253-67
references.pmid: 11991680
references.citation: Mol Pharmacol. 2007 Jul;72(1):29-39
references.pmid: 17395690
references.citation: Apoptosis. 2008 Jan;13(1):1-9
references.pmid: 17990121
references.citation: Int J Mol Sci. 2013;14(1):1370-82
references.pmid: 23344047
references.citation: Mol Med Rep. 2013 May;7(5):1487-93
references.pmid: 23525225
references.citation: J Biol Chem. 2012 Apr 6;287(15):11761-8
references.pmid: 22354961
references.citation: J Pharmacol Sci. 2007 Dec;105(4):317-25
references.pmid: 18094523
references.citation: Int J Biochem Cell Biol. 2004 Dec;36(12):2445-62
references.pmid: 15325584
references.citation: Int J Biochem Cell Biol. 2009 Nov;41(11):2334-42
references.pmid: 19563910
references.citation: Mol Med Rep. 2010 Jan-Feb;3(1):63-7
references.pmid: 21472201
references.citation: Chem Biol Interact. 2013 Nov 25;206(2):279-88
references.pmid: 24121004
references.citation: Nature. 2001 Mar 1;410(6824):37-40
references.pmid: 11242034
references.citation: EMBO J. 2000 Nov 1;19(21):5720-8
references.pmid: 11060023
references.citation: Cancer Lett. 2014 Feb 28;343(2):239-48
references.pmid: 24099910
references.citation: Autophagy. 2009 Aug;5(6):824-34
references.pmid: 19556857
references.citation: Oncogene. 2007 May 14;26(22):3100-12
references.pmid: 17496909
references.citation: Int J Mol Med. 2013 Jul;32(1):85-92
references.pmid: 23652631
references.citation: Int J Mol Sci. 2014;15(2):3145-53
references.pmid: 24566140
references.citation: Toxicol Lett. 2012 Apr 5;210(1):78-86
references.pmid: 22322152
references.citation: Nat Rev Cancer. 2005 Sep;5(9):726-34
references.pmid: 16148885
references.citation: Cell Death Differ. 2005 Nov;12 Suppl 2:1542-52
references.pmid: 16247502
references.citation: Cancer Lett. 2012 Jul 28;320(2):171-9
references.pmid: 22406827
references.citation: Oncotarget. 2012 Apr;3(4):371-94
references.pmid: 22564882
references.citation: Autophagy. 2008 Oct;4(7):949-51
references.pmid: 18769111
references.citation: Crit Rev Eukaryot Gene Expr. 2011;21(1):71-100
references.pmid: 21967333
references.citation: J Chromatogr A. 1996 Apr 12;730(1-2):17-23
references.pmid: 8680588
references.citation: Oncol Lett. 2010 May;1(3):489-493
references.pmid: 22966330
references.citation: Nat Rev Clin Oncol. 2011 Sep;8(9):528-39
references.pmid: 21587219
references.citation: Mol Med Rep. 2010 Mar-Apr;3(2):227-31
references.pmid: 21472226
references.citation: Biol Pharm Bull. 2006 Dec;29(12):2388-94
references.pmid: 17142969
references.citation: Carcinogenesis. 2013 Jun;34(6):1331-42
references.pmid: 23393227
references.citation: Dev Cell. 2004 Apr;6(4):463-77
references.pmid: 15068787