Long-term ethanol exposure causes human liver cancer cells to become resistant to mitomycin c treatment through the inactivation of bad-mediated apoptosis

Ching Shui Huang, Yi Ru Lee, Ching Shyang Chen, Shih Hsin Tu, Ying-Jan Wang, Chia Hwa Lee, Li Ching Chen, Hui Wen Chang, Chien Hsi Chang, Su Chih-Ming, Chih Hsiung Wu, Yuan Soon Ho

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

The aim of this study was to test whether long-term ethanol consumption confers therapeutic resistance to human liver cancer patients infected with hepatitis B virus (HBV). Chronic ethanol-treated cells were established by consecutively culturing a human hepatocellular carcinoma cell line, Hep 3B, which contains integrated HBV sequences, for 20-40 passages with or without 10mM ethanol (designated as E20-E40 and C20-C40, respectively). Flow cytometry analysis demonstrated that a growth promoting effect of long-term ethanol treatment was induced in the E40 cells through preferential acceleration of S-phase in these cells. Lower protein expression levels of p16, p21/Cip1, and p27/Kip1 were detected in the ethanol-treated E40 cells. We further demonstrated that long-term ethanol-treated E40 cells develop drug resistance in response to mitomycin C (MMC) treatment (>8 μM). Immunoblot analysis revealed that caspase-8-mediated mitochondrial apoptotic signals (such as Bad) were inactivated in the MMC-resistant E40 cells. Immunoprecipitation experiments demonstrated that the sequestration of phosphorylated Bad (Ser-112) through its binding with 14-3-3 was detected more profoundly in the MMC-resistant E40 cells. Next, we examined the therapeutic efficacy of MMC (10mg MMC/kg body weight, three times per week) in severe combined immunodeficient (SCID) mice bearing E40- and C40-xenografted tumors. Significant reductions (>3-fold) in tumor growth were detected in MMC-treated C40-xenografted mice. In vivo and in vitro studies demonstrated that AKT-and extracellular signal-regulated kinase (ERK)-mediated survival factors inhibited the Bad-induced mitochondrial apoptotic signals that were involved in E40 tumor cells and that conferred resistance to MMC.

Original languageEnglish
Pages (from-to)728-738
Number of pages11
JournalMolecular Carcinogenesis
Volume49
Issue number8
DOIs
Publication statusPublished - 2010 Aug 1

Fingerprint

Mitomycin
Liver Neoplasms
Ethanol
Apoptosis
Therapeutics
Hepatitis B virus
Neoplasms
SCID Mice
Caspase 8
Extracellular Signal-Regulated MAP Kinases
Growth
S Phase
Immunoprecipitation
Drug Resistance
Hepatocellular Carcinoma
Flow Cytometry
Body Weight
Cell Line
Survival

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cancer Research

Cite this

Huang, Ching Shui ; Lee, Yi Ru ; Chen, Ching Shyang ; Tu, Shih Hsin ; Wang, Ying-Jan ; Lee, Chia Hwa ; Chen, Li Ching ; Chang, Hui Wen ; Chang, Chien Hsi ; Chih-Ming, Su ; Wu, Chih Hsiung ; Ho, Yuan Soon. / Long-term ethanol exposure causes human liver cancer cells to become resistant to mitomycin c treatment through the inactivation of bad-mediated apoptosis. In: Molecular Carcinogenesis. 2010 ; Vol. 49, No. 8. pp. 728-738.
@article{5b5c7948c0b449f3b5c07db496f15e62,
title = "Long-term ethanol exposure causes human liver cancer cells to become resistant to mitomycin c treatment through the inactivation of bad-mediated apoptosis",
abstract = "The aim of this study was to test whether long-term ethanol consumption confers therapeutic resistance to human liver cancer patients infected with hepatitis B virus (HBV). Chronic ethanol-treated cells were established by consecutively culturing a human hepatocellular carcinoma cell line, Hep 3B, which contains integrated HBV sequences, for 20-40 passages with or without 10mM ethanol (designated as E20-E40 and C20-C40, respectively). Flow cytometry analysis demonstrated that a growth promoting effect of long-term ethanol treatment was induced in the E40 cells through preferential acceleration of S-phase in these cells. Lower protein expression levels of p16, p21/Cip1, and p27/Kip1 were detected in the ethanol-treated E40 cells. We further demonstrated that long-term ethanol-treated E40 cells develop drug resistance in response to mitomycin C (MMC) treatment (>8 μM). Immunoblot analysis revealed that caspase-8-mediated mitochondrial apoptotic signals (such as Bad) were inactivated in the MMC-resistant E40 cells. Immunoprecipitation experiments demonstrated that the sequestration of phosphorylated Bad (Ser-112) through its binding with 14-3-3 was detected more profoundly in the MMC-resistant E40 cells. Next, we examined the therapeutic efficacy of MMC (10mg MMC/kg body weight, three times per week) in severe combined immunodeficient (SCID) mice bearing E40- and C40-xenografted tumors. Significant reductions (>3-fold) in tumor growth were detected in MMC-treated C40-xenografted mice. In vivo and in vitro studies demonstrated that AKT-and extracellular signal-regulated kinase (ERK)-mediated survival factors inhibited the Bad-induced mitochondrial apoptotic signals that were involved in E40 tumor cells and that conferred resistance to MMC.",
author = "Huang, {Ching Shui} and Lee, {Yi Ru} and Chen, {Ching Shyang} and Tu, {Shih Hsin} and Ying-Jan Wang and Lee, {Chia Hwa} and Chen, {Li Ching} and Chang, {Hui Wen} and Chang, {Chien Hsi} and Su Chih-Ming and Wu, {Chih Hsiung} and Ho, {Yuan Soon}",
year = "2010",
month = "8",
day = "1",
doi = "10.1002/mc.20648",
language = "English",
volume = "49",
pages = "728--738",
journal = "Molecular Carcinogenesis",
issn = "0899-1987",
publisher = "Wiley-Liss Inc.",
number = "8",

}

Huang, CS, Lee, YR, Chen, CS, Tu, SH, Wang, Y-J, Lee, CH, Chen, LC, Chang, HW, Chang, CH, Chih-Ming, S, Wu, CH & Ho, YS 2010, 'Long-term ethanol exposure causes human liver cancer cells to become resistant to mitomycin c treatment through the inactivation of bad-mediated apoptosis', Molecular Carcinogenesis, vol. 49, no. 8, pp. 728-738. https://doi.org/10.1002/mc.20648

Long-term ethanol exposure causes human liver cancer cells to become resistant to mitomycin c treatment through the inactivation of bad-mediated apoptosis. / Huang, Ching Shui; Lee, Yi Ru; Chen, Ching Shyang; Tu, Shih Hsin; Wang, Ying-Jan; Lee, Chia Hwa; Chen, Li Ching; Chang, Hui Wen; Chang, Chien Hsi; Chih-Ming, Su; Wu, Chih Hsiung; Ho, Yuan Soon.

In: Molecular Carcinogenesis, Vol. 49, No. 8, 01.08.2010, p. 728-738.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Long-term ethanol exposure causes human liver cancer cells to become resistant to mitomycin c treatment through the inactivation of bad-mediated apoptosis

AU - Huang, Ching Shui

AU - Lee, Yi Ru

AU - Chen, Ching Shyang

AU - Tu, Shih Hsin

AU - Wang, Ying-Jan

AU - Lee, Chia Hwa

AU - Chen, Li Ching

AU - Chang, Hui Wen

AU - Chang, Chien Hsi

AU - Chih-Ming, Su

AU - Wu, Chih Hsiung

AU - Ho, Yuan Soon

PY - 2010/8/1

Y1 - 2010/8/1

N2 - The aim of this study was to test whether long-term ethanol consumption confers therapeutic resistance to human liver cancer patients infected with hepatitis B virus (HBV). Chronic ethanol-treated cells were established by consecutively culturing a human hepatocellular carcinoma cell line, Hep 3B, which contains integrated HBV sequences, for 20-40 passages with or without 10mM ethanol (designated as E20-E40 and C20-C40, respectively). Flow cytometry analysis demonstrated that a growth promoting effect of long-term ethanol treatment was induced in the E40 cells through preferential acceleration of S-phase in these cells. Lower protein expression levels of p16, p21/Cip1, and p27/Kip1 were detected in the ethanol-treated E40 cells. We further demonstrated that long-term ethanol-treated E40 cells develop drug resistance in response to mitomycin C (MMC) treatment (>8 μM). Immunoblot analysis revealed that caspase-8-mediated mitochondrial apoptotic signals (such as Bad) were inactivated in the MMC-resistant E40 cells. Immunoprecipitation experiments demonstrated that the sequestration of phosphorylated Bad (Ser-112) through its binding with 14-3-3 was detected more profoundly in the MMC-resistant E40 cells. Next, we examined the therapeutic efficacy of MMC (10mg MMC/kg body weight, three times per week) in severe combined immunodeficient (SCID) mice bearing E40- and C40-xenografted tumors. Significant reductions (>3-fold) in tumor growth were detected in MMC-treated C40-xenografted mice. In vivo and in vitro studies demonstrated that AKT-and extracellular signal-regulated kinase (ERK)-mediated survival factors inhibited the Bad-induced mitochondrial apoptotic signals that were involved in E40 tumor cells and that conferred resistance to MMC.

AB - The aim of this study was to test whether long-term ethanol consumption confers therapeutic resistance to human liver cancer patients infected with hepatitis B virus (HBV). Chronic ethanol-treated cells were established by consecutively culturing a human hepatocellular carcinoma cell line, Hep 3B, which contains integrated HBV sequences, for 20-40 passages with or without 10mM ethanol (designated as E20-E40 and C20-C40, respectively). Flow cytometry analysis demonstrated that a growth promoting effect of long-term ethanol treatment was induced in the E40 cells through preferential acceleration of S-phase in these cells. Lower protein expression levels of p16, p21/Cip1, and p27/Kip1 were detected in the ethanol-treated E40 cells. We further demonstrated that long-term ethanol-treated E40 cells develop drug resistance in response to mitomycin C (MMC) treatment (>8 μM). Immunoblot analysis revealed that caspase-8-mediated mitochondrial apoptotic signals (such as Bad) were inactivated in the MMC-resistant E40 cells. Immunoprecipitation experiments demonstrated that the sequestration of phosphorylated Bad (Ser-112) through its binding with 14-3-3 was detected more profoundly in the MMC-resistant E40 cells. Next, we examined the therapeutic efficacy of MMC (10mg MMC/kg body weight, three times per week) in severe combined immunodeficient (SCID) mice bearing E40- and C40-xenografted tumors. Significant reductions (>3-fold) in tumor growth were detected in MMC-treated C40-xenografted mice. In vivo and in vitro studies demonstrated that AKT-and extracellular signal-regulated kinase (ERK)-mediated survival factors inhibited the Bad-induced mitochondrial apoptotic signals that were involved in E40 tumor cells and that conferred resistance to MMC.

UR - http://www.scopus.com/inward/record.url?scp=77956135374&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77956135374&partnerID=8YFLogxK

U2 - 10.1002/mc.20648

DO - 10.1002/mc.20648

M3 - Article

VL - 49

SP - 728

EP - 738

JO - Molecular Carcinogenesis

JF - Molecular Carcinogenesis

SN - 0899-1987

IS - 8

ER -