title: Inhibitory effect of Curcuma purpurascens BI. rhizome on HT-29 colon cancer cells through mitochondrial-dependent apoptosis pathway.
abstract: BACKGROUND
  Curcuma purpurascens BI. (Zingiberaceae) commonly known as 'Koneng Tinggang' and 'Temu Tis' is a Javanese medicinal plant which has been used for numerous ailments and diseases in rural Javanese communities. In the present study, the apoptogenic activity of dichloromethane extract of Curcuma purpurascens BI. rhizome (DECPR) was investigated against HT-29 human colon cancer cells.
  
  
  METHODS
  Acute toxicity study of DECPR was performed in Sprague-Dawley rats. Compounds of DECPR were analyzed by the gas chromatography-mass spectrometry-time of flight (GC-MS-TOF) analysis. Cytotoxic effect of DECPR on HT-29 cells was analyzed by MTT and lactate dehydrogenase (LDH) assays. Effects of DECPR on reactive oxygen species (ROS) formation and mitochondrial-initiated events were investigated using a high content screening system. The activities of the caspases were also measured using a fluorometric assay. The quantitative PCR analysis was carried out to examine the gene expression of Bax, Bcl-2 and Bcl-xl proteins.
  
  
  RESULTS
  The in vivo acute toxicity study of DECPR on rats showed the safety of this extract at the highest dose of 5 g/kg. The GC-MS-TOF analysis of DECPR detected turmerone as the major compound in dichloromethane extract. IC50 value of DECPR towards HT-29 cells after 24 h treatment was found to be 7.79 ± 0.54 μg/mL. In addition, DECPR induced LDH release and ROS generation in HT-29 cells through a mechanism involving nuclear fragmentation and cytoskeletal rearrangement. The mitochondrial-initiated events, including collapse in mitochondrial membrane potential and cytochrome c leakage was also triggered by DECPR treatment. Initiator caspase-9 and executioner caspase-3 was dose-dependently activated by DECPR. The quantitative PCR analysis on the mRNA expression of Bcl-2 family of proteins showed a significant up-regulation of Bax associated with down-regulation in Bcl-2 and Bcl-xl mRNA expression.
  
  
  CONCLUSIONS
  The findings presented in the current study showed that DECP suppressed the proliferation of HT-29 colon cancer cells and triggered the induction of apoptosis through mitochondrial-dependent pathway.
authors: Department of Pharmacology, Faculty of Medicine, University of Malaya, 50603, Kuala Lumpur, Malaysia. e_rouhollahi@yahoo.com.
authors: Elham
authors: E
authors: Rouhollahi
authors: Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603, Kuala Lumpur, Malaysia. soheil.zorofchian@gmail.com.
authors: Soheil
authors: S
authors: Zorofchian Moghadamtousi
authors: Department of Pharmacology, Faculty of Medicine, University of Malaya, 50603, Kuala Lumpur, Malaysia. paydarmj@gmail.com.
authors: Mohammadjavad
authors: M
authors: Paydar
authors: Department of Chemistry, Faculty of Science, University of Malaya, 50603, Kuala Lumpur, Malaysia. mehranfadaie_n@yahoo.com.
authors: Mehran
authors: M
authors: Fadaeinasab
authors: Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603, Kuala Lumpur, Malaysia. z_maryam81@yahoo.com.
authors: Maryam
authors: M
authors: Zahedifard
authors: Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603, Kuala Lumpur, Malaysia. maryam_hajrezaie@yahoo.com.
authors: Maryam
authors: M
authors: Hajrezaie
authors: Department of Chemistry, Faculty of Science, University of Malaya, 50603, Kuala Lumpur, Malaysia. omerhamdi2001@hotmail.com.
authors: Omer Abdalla
authors: OA
authors: Ahmed Hamdi
authors: Department of Pharmacology, Faculty of Medicine, University of Malaya, 50603, Kuala Lumpur, Malaysia. looicy@um.edu.my.
authors: Chung Yeng
authors: CY
authors: Looi
authors: Department of Biomedical Science, Faculty of Medicine, University of Malaya, 50603, Kuala Lumpur, Malaysia. ammeen@um.edu.my.
authors: Mahmood Ameen
authors: MA
authors: Abdulla
authors: Department of Chemistry, Faculty of Science, University of Malaya, 50603, Kuala Lumpur, Malaysia. khalijah@um.edu.my.
authors: Khalijah
authors: K
authors: Awang
authors: Department of Pharmacology, Faculty of Medicine, University of Malaya, 50603, Kuala Lumpur, Malaysia. zahurin@um.edu.my.
authors: Zahurin
authors: Z
authors: Mohamed
mesh_terms: D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology / Q000627:therapeutic use*; D017209:Apoptosis / Q000187:drug effects; D020169:Caspases / Q000378:metabolism; D003110:Colonic Neoplasms / Q000188:drug therapy* / Q000378:metabolism; D030024:Curcuma / Q000737:chemistry*; D045304:Cytochromes c / Q000378:metabolism; D019073:HT29 Cells; D006801:Humans; D008297:Male; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects*; D008928:Mitochondria / Q000187:drug effects*; D008517:Phytotherapy; D010936:Plant Extracts / Q000737:chemistry / Q000494:pharmacology / Q000627:therapeutic use*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000378:metabolism; D027343:Rhizome; D015398:Signal Transduction / Q000187:drug effects; D030521:Zingiberaceae; D051028:bcl-2-Associated X Protein / Q000378:metabolism
chemical_list: D000972:Antineoplastic Agents, Phytogenic; C494828:Bax protein, rat; D010936:Plant Extracts; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D051028:bcl-2-Associated X Protein; D045304:Cytochromes c; D020169:Caspases
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