title: Oldenlandia diffusa suppresses metastatic potential through inhibiting matrix metalloproteinase-9 and intercellular adhesion molecule-1 expression via p38 and ERK1/2 MAPK pathways and induces apoptosis in human breast cancer MCF-7 cells.
abstract: BACKGROUND
  Oldenlandia diffusa (OD) has long been known as an apoptotic inducer in breast tumors in ethnomedicine.
  
  
  OBJECTIVE
  To scientifically confirm the anti-breast cancer effects of water, methanol (MeOH) and butanol (BuOH) extracts of O. diffusa on cell apoptosis, matrix metalloproteinases (MMPs), intercellular adhesion molecule (ICAM)-1 and intracellular signaling in MCF-7 breast cancer cells.
  
  
  METHODS
  MeOH extracts (MOD) and BuOH extracts (BOD) were prepared and examined for their ability to inhibit phorbol myristate acetate (PMA)-induced matrix metalloproteinase (MMP)-9 and intercellular adhesion molecule (ICAM)-1 expressions in MCF-7 human breast cancer cells. Additionally, transwell migration, invasion and transcriptional activity were assessed. Results of immunofluorescence confocal microscopy for translocation of NF-κB and p-ERK and p-p38 were also checked. Finally, apoptotic signals including processed caspase-8, caspase-7, poly ADP-ribose polymerase, Bax and Bcl-2 were examined.
  
  
  RESULTS
  MOD and BOD specifically inhibited PMA-induced MMP-9 expression as well as invasive and migration potential via ICAM-1. The inhibitory activity was also based on the suppressed transcriptional activity in MCF-7 breast cancer cells. Results of immunofluorescence confocal microscopy showed that translocation of NF-κB decreased upon BOD and MOD treatments, with a decreased level of p-ERK and p-p38 phosphorylation. In addition, treatment of MCF-7 cells with MOD and BOD activated apoptosis-linked proteins including enzymatically active forms of processed caspase-8, caspase-7 and poly ADP-ribose polymerase, together with increased expression of mitochondrial apoptotic protein, Bax and decreased expression of Bcl-2.
  
  
  CONCLUSIONS
  The results indicate that OD as an anti-metastatic agent suppresses the metastatic response by targeting p-ERK, p-38 and NF-κB, thus reducing the invasion capacity of MCF-7 breast cancer cells through inhibition of MMP-9 and ICAM-1 expression and plays an important role in the regulation of breast cancer cell apoptosis.
authors.affiliation: Division of Applied Medicine, School of Korean Medicine and Research Center for Healthy Aging, Pusan National University, Yangsan City, Gyeongsangnam-Do, Korea. Electronic address: chungtw@hanmail.net.
authors.forename: Tae-Wook
authors.initials: TW
authors.lastname: Chung
authors.affiliation: Department of Biological Sciences, SungKyunKwan University, Seoburo 2066, Jangan-Gu, Suwon, Gyunggi-Do 16419, Korea. Electronic address: chj204@skku.edu.
authors.forename: Hyunju
authors.initials: H
authors.lastname: Choi
authors.affiliation: Department of Biological Sciences, SungKyunKwan University, Seoburo 2066, Jangan-Gu, Suwon, Gyunggi-Do 16419, Korea. Electronic address: jml@skku.edu.
authors.forename: Ji-Min
authors.initials: JM
authors.lastname: Lee
authors.affiliation: Department of Biological Sciences, SungKyunKwan University, Seoburo 2066, Jangan-Gu, Suwon, Gyunggi-Do 16419, Korea. Electronic address: sunghyungha@skku.edu.
authors.forename: Sun-Hyung
authors.initials: SH
authors.lastname: Ha
authors.affiliation: Department of Biological Sciences, SungKyunKwan University, Seoburo 2066, Jangan-Gu, Suwon, Gyunggi-Do 16419, Korea. Electronic address: kwak@skku.edu.
authors.forename: Choong-Hwan
authors.initials: CH
authors.lastname: Kwak
authors.affiliation: Department of Biological Sciences, SungKyunKwan University, Seoburo 2066, Jangan-Gu, Suwon, Gyunggi-Do 16419, Korea. Electronic address: pokusa6@hotmail.com.
authors.forename: Fukushi
authors.initials: F
authors.lastname: Abekura
authors.affiliation: Department of Biological Sciences, SungKyunKwan University, Seoburo 2066, Jangan-Gu, Suwon, Gyunggi-Do 16419, Korea. Electronic address: wnsdud2057@naver.com.
authors.forename: Jun-Young
authors.initials: JY
authors.lastname: Park
authors.affiliation: Research Institute of Biomedical Engineering and Department of Medicine, Catholic University of Daegu School of Medicine, Daegu, Korea. Electronic address: ycchang@cu.ac.kr.
authors.forename: Young-Chae
authors.initials: YC
authors.lastname: Chang
authors.affiliation: Division of Applied Medicine, School of Korean Medicine and Research Center for Healthy Aging, Pusan National University, Yangsan City, Gyeongsangnam-Do, Korea. Electronic address: haggis@pnu.ac.kr.
authors.forename: Ki-Tae
authors.initials: KT
authors.lastname: Ha
authors.affiliation: Division of Enteric Diseases, Center for Infectious Diseases Research, Korea National Institute of Health, Heungdeok-gu, Cheongju 363-951, Korea. Electronic address: skcho38@hotmail.com.
authors.forename: Seung-Hak
authors.initials: SH
authors.lastname: Cho
authors.affiliation: College of Pharmacy, Yeungnam University, Gyeongsan 712-749, Korea. Electronic address: hwchang@yu.ac.kr.
authors.forename: Hyeun Wook
authors.initials: HW
authors.lastname: Chang
authors.affiliation: Department of Medicinal Biotechnology, College of Health Sciences, Dong-A University, Busan 49315, Korea. Electronic address: yclee@dau.ac.kr.
authors.forename: Young-Choon
authors.initials: YC
authors.lastname: Lee
authors.affiliation: Division of Applied Medicine, School of Korean Medicine and Research Center for Healthy Aging, Pusan National University, Yangsan City, Gyeongsangnam-Do, Korea. Electronic address: chkimbio@skku.edu.
authors.forename: Cheorl-Ho
authors.initials: CH
authors.lastname: Kim
mesh_terms: D000972:Antineoplastic Agents, Phytogenic / Q000302:isolation & purification / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism; D001943:Breast Neoplasms / Q000188:drug therapy* / Q000201:enzymology / Q000235:genetics / Q000473:pathology; D000440:Butanols / Q000737:chemistry; D002465:Cell Movement / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000235:genetics / Q000378:metabolism*; D061986:MCF-7 Cells; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism*; D000432:Methanol / Q000737:chemistry; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism*; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism*; D016328:NF-kappa B / Q000378:metabolism; D009361:Neoplasm Invasiveness; D009362:Neoplasm Metastasis; D032067:Oldenlandia / Q000737:chemistry*; D010766:Phosphorylation; D008517:Phytotherapy; D010936:Plant Extracts / Q000302:isolation & purification / Q000494:pharmacology*; D010946:Plants, Medicinal; D015398:Signal Transduction / Q000187:drug effects; D014158:Transcription, Genetic / Q000187:drug effects; D014162:Transfection; D014867:Water / Q000737:chemistry; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism*
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D051017:Apoptosis Regulatory Proteins; D000440:Butanols; C573420:ICAM1 protein, human; D016328:NF-kappa B; D010936:Plant Extracts; D014867:Water; D018799:Intercellular Adhesion Molecule-1; C535150:MAPK1 protein, human; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D048051:p38 Mitogen-Activated Protein Kinases; C579270:MMP9 protein, human; D020780:Matrix Metalloproteinase 9; D000432:Methanol
keywords: Apoptosis; ICAM-1; MCF-7 cells; MMP-9; Oldenlandia diffusa; Oleanolic acid (PubChem CID: 10494); Ssperuloside, E-6-O-p-coumaroyl scandoside methyl ester and E-6-O-p-coumaroyl scandoside methyl ester-10-methyl ether (PubChem CID: 84298); Stigmasterol (PubChem CID: 5280794); Ursolic acid (PubChem CID: 64945)