A natural compound from Hydnophytum formicarium induces apoptosis of MCF-7 cells via up-regulation of BaxReport as inadecuate




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Cancer Cell International

, 10:14

First Online: 04 May 2010Received: 09 July 2009Accepted: 04 May 2010

Abstract

BackgroundHydnophytum formicarium Jack is an epyphytic shrub that belongs to the family of Rubiaceae and is native to the tropical rain forests of the Asean region, which includes Malaysia. A flavanoid derivative, 7, 3-, 5-trihydroxyflavanone 3HFD, isolated from H. formicarium has been reported to have cytotoxic effects on the human breast carcinoma cell line MCF-7. The aim of the current study was to investigate the mode of cell death in MCF-7 cells treated with 3HFD. A DNA fragmentation assay was conducted on isolated genomic DNA, a TUNEL assay was used to determine the mode of cell death and Western blotting was used to evaluate the expression levels of Bax and Bcl-2. Immunofluorescence staining of MCF-7 cells was also performed to confirm the up-regulation of the Bax protein.

ResultsThe ladder pattern resulting from the DNA fragmentation assay was a multimer of 180 kb. The morphological changes of cells undergoing apoptosis were visualised by a TUNEL assay over time. The percentage of apoptotic cells increased as early as 6 hours post treatment compared to untreated cells. Western blotting revealed up-regulation of the pro-apoptotic protein Bax. However, 3HFD did not affect expression of the anti-apoptotic protein Bcl-2.

ConclusionsOur results provide evidence that plant-derived 3HFD was able to induce the apoptotic cell death of MCF-7 cells by increasing Bax expression level and makes 3HFD a promising agent for chemotherapy, which merits further study.

List of abbreviationsATCCAmerican Type Cell Culture Collection

BaxBcl-2-associated × protein

Bcl-2B-cell lymphoma-2

Cacalcium ion

Chang liver cellsnormal liver cells

CO2carbon dioxide

DMEMDulbecco-s modified Eagle-s medium

DMSOdimethylsulfoxide

DNAdeoxyribonucleic acid

dUTPdeoxyuridine triphosphate

ELISAEnzyme Linked Immuno Sorbent Assay

FBSfoetal bovine serum

HClhydrochloride acid

IC50inhibition concentration to kill 50% of cells population

IgGImmunoglobulin-G

MDBK cellsMadin Darby Bovine Kidney cells

PBSphosphate-buffered saline

PVDFpolyvinyl-difluoride

SDSsodium dodecyl sulphate

SSCsodium chloride-sodium citrate

TdTTerminal Deoxynucleotidyl Transferase

TUNELTdT-mediated dUTP nick-end labelling

hhour

ggram

bpbase pair.

Electronic supplementary materialThe online version of this article doi:10.1186-1475-2867-10-14 contains supplementary material, which is available to authorized users.

Judit Hohmann contributed equally to this work.

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Author: Hasmah Abdullah - Azimahtol Hawariah Lope Pihie - Judit Hohmann - Joseph Molnár

Source: https://link.springer.com/



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