title: Anti-cancer effect and the underlying mechanisms of gypenosides on human colorectal cancer SW-480 cells.
abstract: BACKGROUND
  Gypenosides (Gyp), the main components from Gynostemma pentaphyllum Makino, are widely used in traditional Chinese medicine. The present study aimed to investigate the anti-cancer effect and the underlying mechanisms of Gyp on human colorectal cancer cells SW-480.
  
  
  METHODS
  The inhibitory effect of Gyp on SW-480 cells was evaluated by MTT assay. Apoptotic cell death was detected by nuclear Hoechst 33342 staining and DNA fragmentation analysis. Apoptosis was analyzed using Annexin V-PE/7-amino-actinomycin D staining. Cell membrane integrity was evaluated with flow cytometry following PI staining. Changes of mitochondrial membrane potential (Δψm) were detected through flow cytometry analysis of rhodamine 123 (Rh123). The role of reactive oxygen species (ROS) in Gyp induced cell death was investigated by intracellular ROS generation and general ROS scavenger. Wound-healing assay was carried out to investigate Gyp-inhibited migration of SW-480 cells in vitro. Additionally, the alterations in F-actin microfilaments were analyzed by FITC-labeled phalloidin toxin staining and the morphological changes were evaluated under scanning electron microscope (SEM).
  
  
  RESULTS
  After the Gyp treatment, the plasma membrane permeability of SW-480 cell was increased, Δψm was decreased significantly, the level of intracellular ROS level was increased, DNA fragmentation and apoptotic morphology were observed. Cells treated with Gyp exert serious microfilament network collapse as well as the significant decrease in the number of microvilli. Gyp induced the changes of cell viability, cell migration, intracellular ROS generation and nuclear morphology were alleviated obviously by NAC.
  
  
  CONCLUSIONS
  The results in this study implied that ROS play an important role in Gyp induced cell toxicity and apoptosis, and the mitochondria damage may be upstream of ROS generation post Gyp treatment. The findings of the present study provide new evidences for anti-tumor mechanisms by which Gyp induces apoptosis in vitro.
authors.affiliation: Key Laboratory of Medicinal Resources and Natural Pharmaceutical Chemistry, Ministry of Education, National Engineering Laboratory for Resource Developing of Endangered Chinese Crude Drugs in Northwest of China, College of Life Sciences, Shaanxi Normal University, Xi'an, Shaanxi, China.
authors.forename: Han
authors.initials: H
authors.lastname: Yan
authors.affiliation: Key Laboratory of Medicinal Resources and Natural Pharmaceutical Chemistry, Ministry of Education, National Engineering Laboratory for Resource Developing of Endangered Chinese Crude Drugs in Northwest of China, College of Life Sciences, Shaanxi Normal University, Xi'an, Shaanxi, China.
authors.forename: Xiaobing
authors.initials: X
authors.lastname: Wang
authors.affiliation: Key Laboratory of Medicinal Resources and Natural Pharmaceutical Chemistry, Ministry of Education, National Engineering Laboratory for Resource Developing of Endangered Chinese Crude Drugs in Northwest of China, College of Life Sciences, Shaanxi Normal University, Xi'an, Shaanxi, China.
authors.forename: Junfeng
authors.initials: J
authors.lastname: Niu
authors.affiliation: Key Laboratory of Medicinal Resources and Natural Pharmaceutical Chemistry, Ministry of Education, National Engineering Laboratory for Resource Developing of Endangered Chinese Crude Drugs in Northwest of China, College of Life Sciences, Shaanxi Normal University, Xi'an, Shaanxi, China.
authors.forename: Yaqin
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Key Laboratory of Medicinal Resources and Natural Pharmaceutical Chemistry, Ministry of Education, National Engineering Laboratory for Resource Developing of Endangered Chinese Crude Drugs in Northwest of China, College of Life Sciences, Shaanxi Normal University, Xi'an, Shaanxi, China.
authors.forename: Pan
authors.initials: P
authors.lastname: Wang
authors.affiliation: Key Laboratory of Medicinal Resources and Natural Pharmaceutical Chemistry, Ministry of Education, National Engineering Laboratory for Resource Developing of Endangered Chinese Crude Drugs in Northwest of China, College of Life Sciences, Shaanxi Normal University, Xi'an, Shaanxi, China.
authors.forename: Quanhong
authors.initials: Q
authors.lastname: Liu
mesh_terms: D000111:Acetylcysteine / Q000494:pharmacology; D008841:Actin Cytoskeleton / Q000187:drug effects / Q000378:metabolism; D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology* / Q000627:therapeutic use*; D017209:Apoptosis / Q000187:drug effects; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002465:Cell Movement / Q000187:drug effects; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D048430:Cell Shape / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D015179:Colorectal Neoplasms / Q000188:drug therapy* / Q000473:pathology* / Q000648:ultrastructure; D053938:DNA Fragmentation / Q000187:drug effects; D004354:Drug Screening Assays, Antitumor; D005434:Flow Cytometry; D029774:Gynostemma / Q000737:chemistry; D006801:Humans; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D010936:Plant Extracts / Q000737:chemistry / Q000494:pharmacology / Q000627:therapeutic use; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D000970:Antineoplastic Agents; D010936:Plant Extracts; D017382:Reactive Oxygen Species; C474918:gypenoside; D000111:Acetylcysteine
references.citation: Cancer Res. 2006 May 15;66(10):5379-86
references.pmid: 16707465
references.citation: Zhongguo Zhong Xi Yi Jie He Za Zhi. 1993 May;13(5):278-80, 261
references.pmid: 8219678
references.citation: Curr Opin Cell Biol. 1997 Feb;9(1):76-85
references.pmid: 9013677
references.citation: Cancer Res. 1976 Dec;36(12):4562-9
references.pmid: 1000501
references.citation: Apoptosis. 2005 Jan;10(1):201-8
references.pmid: 15711936
references.citation: Endocr Relat Cancer. 1999 Mar;6(1):13-6
references.pmid: 10732780
references.citation: Int Rev Cytol. 2001;202:35-98
references.pmid: 11061563
references.citation: Cancer Biother. 1993 Fall;8(3):263-72
references.pmid: 7804367
references.citation: Cancer Lett. 2008 Sep 28;269(1):7-17
references.pmid: 18462867
references.citation: Eur J Cancer. 2010 Mar;46(4):765-81
references.pmid: 20116997
references.citation: Cancer. 1994 Apr 15;73(8):2013-26
references.pmid: 8156506
references.citation: J Ethnopharmacol. 2005 Aug 22;100(1-2):72-9
references.pmid: 16009521
references.citation: Ceska Slov Farm. 1995 Oct;44(5):246-51
references.pmid: 8520921
references.citation: Cell Biol Int. 2007 Nov;31(11):1353-8
references.pmid: 17587607
references.citation: Lancet. 2005 Jan 8-14;365(9454):153-65
references.pmid: 15639298
references.citation: Chem Biol Interact. 2008 Jan 10;171(1):45-56
references.pmid: 17935706
references.citation: J Ethnopharmacol. 2010 Aug 19;131(1):1-9
references.pmid: 20420897
references.citation: Biochem Pharmacol. 2005 Jul 15;70(2):229-41
references.pmid: 15941563
references.citation: Biochem Biophys Res Commun. 2007 Aug 17;360(1):109-14
references.pmid: 17585878
references.citation: Nitric Oxide. 1999 Oct;3(5):359-65
references.pmid: 10534439
references.citation: J Ethnopharmacol. 1995 May;46(2):125-9
references.pmid: 7650951
references.citation: Am J Clin Nutr. 2009 May;89(5):1441-52
references.pmid: 19339391
references.citation: Hum Exp Toxicol. 2014 May;33(5):522-33
references.pmid: 23900306
references.citation: Cancer Lett. 2006 Feb 20;233(1):28-35
references.pmid: 15899547
references.citation: J Ethnopharmacol. 2012 Nov 21;144(2):322-7
references.pmid: 23000169
references.citation: Cancer Lett. 2002 Sep 26;183(2):169-78
references.pmid: 12065092
references.citation: Am J Chin Med. 1993;21(1):59-69
references.pmid: 8328423
references.citation: Hum Exp Toxicol. 2011 May;30(5):406-15
references.pmid: 20511288
references.citation: Artif Life. 2006 Fall;12(4):617-34
references.pmid: 16953788
references.citation: J Ethnopharmacol. 2011 Sep 2;137(2):1003-10
references.pmid: 21782919
references.citation: Methods. 2008 Mar;44(3):205-21
references.pmid: 18314051
references.citation: Annu Rev Physiol. 1991;53:585-605
references.pmid: 2042972
references.citation: Biochem Pharmacol. 2008 Jul 1;76(1):11-8
references.pmid: 18462707
references.citation: Cancer Chemother Pharmacol. 2005 Jul;56(1):29-38
references.pmid: 15726366
references.citation: Integr Cancer Ther. 2011 Mar;10(1):101-12
references.pmid: 20702487
references.citation: Ann Nutr Metab. 2009;55(1-3):140-61
references.pmid: 19752540
references.citation: Am J Chin Med. 2000;28(1):87-96
references.pmid: 10794120
references.citation: J Ethnopharmacol. 2012 Aug 30;143(1):49-56
references.pmid: 22687253