Limits...
Ethyl gallate induces apoptosis of HL-60 cells by promoting the expression of caspases-8, -9, -3, apoptosis-inducing factor and endonuclease G.

Kim WH, Song HO, Choi HJ, Bang HI, Choi DY, Park H - Int J Mol Sci (2012)

Bottom Line: Western blotting analysis indicated that EG-mediated HL-60 apoptosis mainly occurred through the mitochondrial pathway, as shown by the release of cytochrome c, apoptosis-inducing factor (AIF), and endonuclease G (Endo G), as well as the upregulation of Bcl-2-associated X protein (Bax).EG also activated the death receptor-dependent pathway of apoptosis by enhancing the expression of caspases-8, -9, and -3 and the Bcl-2 interacting domain (Bid).Collectively, our results showed that EG induces apoptosis in HL-60 via mitochondrial-mediated pathways.

View Article: PubMed Central - PubMed

Affiliation: Department of Infection Biology, Zoonosis Research Center, Wonkwang University School of Medicine, 344-2, Shinyong-dong, Iksan, Chonbuk 570-749, South Korea; E-Mails: woong621@gmail.com (W.-H.K.); sea5328@daum.net (H.-O.S.); rerived@empal.com (H.-J.C.).

ABSTRACT
Many phytochemicals have been recognized to have potential therapeutic efficacy in cancer treatment. In this study, we investigated ethyl gallate (EG) for possible proapoptotic effects in the human promyelocytic leukemia cell line, HL-60. We examined cell viability, morphological changes, DNA content and fragmentation, and expression of apoptosis-related proteins for up to 48 h after EG treatment. The results showed that EG induced morphological changes and DNA fragmentation and reduced HL-60 cell viability in a dose-dependent and time-dependent manner. Western blotting analysis indicated that EG-mediated HL-60 apoptosis mainly occurred through the mitochondrial pathway, as shown by the release of cytochrome c, apoptosis-inducing factor (AIF), and endonuclease G (Endo G), as well as the upregulation of Bcl-2-associated X protein (Bax). EG also activated the death receptor-dependent pathway of apoptosis by enhancing the expression of caspases-8, -9, and -3 and the Bcl-2 interacting domain (Bid). Collectively, our results showed that EG induces apoptosis in HL-60 via mitochondrial-mediated pathways.

Show MeSH

Related in: MedlinePlus

Chemical structure of ethyl gallate.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC3472783&req=5

f4-ijms-13-11912: Chemical structure of ethyl gallate.

Mentions: Propidium iodide (PI), dimethyl sulfoxide (DMSO), ribonuclease A (RNase A), and 4,6-diamidino-2-phenylindole dihydrochloride (DAPI) were obtained from Sigma-Aldrich Corp. (St. Louis, MO, USA). Antibodies specific for the following proteins were purchased from the indicated sources. Caspase-3 (Cell Signaling Technology; Danvers, MA, USA); caspase-8, caspase-9, and Endo G (Enzo Life Sciences; Farmingdale, NY, USA); AIF (Bethyl Laboratories; Montgomery, TX, USA); B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), and Bcl-2-interacting domain (Bid) (Santa Cruz Biotechnology; Santa Cruz, CA, USA); cytochrome c (BioVision; Conesa, Buenos Aires, Argentina); and β-actin (Sigma-Aldrich; St. Louis, MO, USA). RPMI 1640 medium, penicillin-streptomycin, fetal bovine serum (FBS), and l-glutamine were obtained from Gibco-BRL (Grand Island, NY, USA). Ethyl gallate isolated from Galla Rhois was obtained from Dr. Youn-Chul Kim’s Laboratory at the College of Pharmacy, Wonkwang University, Iksan, South Korea (Figure 4). All chemicals were of reagent grade.


Ethyl gallate induces apoptosis of HL-60 cells by promoting the expression of caspases-8, -9, -3, apoptosis-inducing factor and endonuclease G.

Kim WH, Song HO, Choi HJ, Bang HI, Choi DY, Park H - Int J Mol Sci (2012)

Chemical structure of ethyl gallate.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC3472783&req=5

f4-ijms-13-11912: Chemical structure of ethyl gallate.
Mentions: Propidium iodide (PI), dimethyl sulfoxide (DMSO), ribonuclease A (RNase A), and 4,6-diamidino-2-phenylindole dihydrochloride (DAPI) were obtained from Sigma-Aldrich Corp. (St. Louis, MO, USA). Antibodies specific for the following proteins were purchased from the indicated sources. Caspase-3 (Cell Signaling Technology; Danvers, MA, USA); caspase-8, caspase-9, and Endo G (Enzo Life Sciences; Farmingdale, NY, USA); AIF (Bethyl Laboratories; Montgomery, TX, USA); B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), and Bcl-2-interacting domain (Bid) (Santa Cruz Biotechnology; Santa Cruz, CA, USA); cytochrome c (BioVision; Conesa, Buenos Aires, Argentina); and β-actin (Sigma-Aldrich; St. Louis, MO, USA). RPMI 1640 medium, penicillin-streptomycin, fetal bovine serum (FBS), and l-glutamine were obtained from Gibco-BRL (Grand Island, NY, USA). Ethyl gallate isolated from Galla Rhois was obtained from Dr. Youn-Chul Kim’s Laboratory at the College of Pharmacy, Wonkwang University, Iksan, South Korea (Figure 4). All chemicals were of reagent grade.

Bottom Line: Western blotting analysis indicated that EG-mediated HL-60 apoptosis mainly occurred through the mitochondrial pathway, as shown by the release of cytochrome c, apoptosis-inducing factor (AIF), and endonuclease G (Endo G), as well as the upregulation of Bcl-2-associated X protein (Bax).EG also activated the death receptor-dependent pathway of apoptosis by enhancing the expression of caspases-8, -9, and -3 and the Bcl-2 interacting domain (Bid).Collectively, our results showed that EG induces apoptosis in HL-60 via mitochondrial-mediated pathways.

View Article: PubMed Central - PubMed

Affiliation: Department of Infection Biology, Zoonosis Research Center, Wonkwang University School of Medicine, 344-2, Shinyong-dong, Iksan, Chonbuk 570-749, South Korea; E-Mails: woong621@gmail.com (W.-H.K.); sea5328@daum.net (H.-O.S.); rerived@empal.com (H.-J.C.).

ABSTRACT
Many phytochemicals have been recognized to have potential therapeutic efficacy in cancer treatment. In this study, we investigated ethyl gallate (EG) for possible proapoptotic effects in the human promyelocytic leukemia cell line, HL-60. We examined cell viability, morphological changes, DNA content and fragmentation, and expression of apoptosis-related proteins for up to 48 h after EG treatment. The results showed that EG induced morphological changes and DNA fragmentation and reduced HL-60 cell viability in a dose-dependent and time-dependent manner. Western blotting analysis indicated that EG-mediated HL-60 apoptosis mainly occurred through the mitochondrial pathway, as shown by the release of cytochrome c, apoptosis-inducing factor (AIF), and endonuclease G (Endo G), as well as the upregulation of Bcl-2-associated X protein (Bax). EG also activated the death receptor-dependent pathway of apoptosis by enhancing the expression of caspases-8, -9, and -3 and the Bcl-2 interacting domain (Bid). Collectively, our results showed that EG induces apoptosis in HL-60 via mitochondrial-mediated pathways.

Show MeSH
Related in: MedlinePlus