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Arsenic trioxide reduces chemo-resistance to 5-fluorouracil and cisplatin in HBx-HepG2 cells via complex mechanisms
BACKGROUND: Multidrug resistance is one of the major reasons chemotherapy-based treatments failed in hepatitis B virus (HBV) related hepatocellular carcinoma (HCC). Hypoxia is generally associated with tumor chemo-resistance. The aim of the study was to investigate the effect of Arsenic trioxide (As...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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BioMed Central
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4676851/ https://www.ncbi.nlm.nih.gov/pubmed/26692822 http://dx.doi.org/10.1186/s12935-015-0269-y |
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author | Yu, Guifang Chen, Xuezhu Chen, Shudi Ye, Weipeng Hou, Kailian Liang, Min |
author_facet | Yu, Guifang Chen, Xuezhu Chen, Shudi Ye, Weipeng Hou, Kailian Liang, Min |
author_sort | Yu, Guifang |
collection | PubMed |
description | BACKGROUND: Multidrug resistance is one of the major reasons chemotherapy-based treatments failed in hepatitis B virus (HBV) related hepatocellular carcinoma (HCC). Hypoxia is generally associated with tumor chemo-resistance. The aim of the study was to investigate the effect of Arsenic trioxide (As(2)O(3)) on the hypoxia-induced chemo-resistance to 5-FU or cisplatin and explored its underlying mechanism in the HBx-HepG2 cells. METHODS: MTT assay was used to examine the cell viability. Mitochondrial membrane potential (MMP) and cell cycle was examined by flow cytometry. qRT-PCR was employed to observe the mRNA expression level; and western blot assay was used to determine the protein expression level. RESULTS: Our results showed that transfection of HBx plasmid established the HBx-HepG2 cells expressing HBx, and the expression of HBx was confirmed by qRT-PCR and western blot. Exposure of HBx-HepG2 cells to hypoxia (5 % O(2), 3 % O(2), 1 % O(2)) for 48 h increased the chemo-resistance to 5-fluorouracil (5-FU) (50–1600 µM) and cisplatin (25–800 µM), reduced MMP, and caused the cell cycle arrest at G(0)/G(1) phase in a concentration-dependent manner. Hypoxia also concentration-dependently (5 % O(2), 3 % O(2), 1 % O(2)) reduced mRNA expression level of P-glycoprotein (P-gp), multidrug resistance protein (MRP1), lung resistance protein (LRP), and decreased the protein expression level of hypoxia-inducible factor-1α (HIF-1α), P-gp MRP1, and LRP. Following pretreatment with As(2)O(3) at a non-cytotoxic concentration re-sensitized the hypoxia (1 % O(2))-induced chemo-resistance to 5-FU and cisplatin in HBx-HepG2 cells. As(2)O(3) pretreatment also prevented MMP reduction and G(0)/G(1) arrest induced by hypoxia. Meanwhile, As(2)O(3) antagonized increase of HIF-1α protein induced by hypoxia, and it also suppresses the increase in expression levels of P-gp, MRP1, and LRP mRNA and proteins. In addition, As(2)O(3) in combination with 5-FU treatment caused up-regulation of DR5, caspase 3, caspase 8, and caspase 9, and down-regulation of BCL-2, but had no effect of DR4. CONCLUSIONS: Our results may suggest that As(2)O(3) re-sensitizes hypoxia-induced chemo-resistance in HBx-HepG2 via complex pathways, and As(2)O(3) may be a potential agent that given in combination with other anti-drugs for the treatment of HBV related HCC, which is resistant to chemotherapy. |
format | Online Article Text |
id | pubmed-4676851 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-46768512015-12-13 Arsenic trioxide reduces chemo-resistance to 5-fluorouracil and cisplatin in HBx-HepG2 cells via complex mechanisms Yu, Guifang Chen, Xuezhu Chen, Shudi Ye, Weipeng Hou, Kailian Liang, Min Cancer Cell Int Primary Research BACKGROUND: Multidrug resistance is one of the major reasons chemotherapy-based treatments failed in hepatitis B virus (HBV) related hepatocellular carcinoma (HCC). Hypoxia is generally associated with tumor chemo-resistance. The aim of the study was to investigate the effect of Arsenic trioxide (As(2)O(3)) on the hypoxia-induced chemo-resistance to 5-FU or cisplatin and explored its underlying mechanism in the HBx-HepG2 cells. METHODS: MTT assay was used to examine the cell viability. Mitochondrial membrane potential (MMP) and cell cycle was examined by flow cytometry. qRT-PCR was employed to observe the mRNA expression level; and western blot assay was used to determine the protein expression level. RESULTS: Our results showed that transfection of HBx plasmid established the HBx-HepG2 cells expressing HBx, and the expression of HBx was confirmed by qRT-PCR and western blot. Exposure of HBx-HepG2 cells to hypoxia (5 % O(2), 3 % O(2), 1 % O(2)) for 48 h increased the chemo-resistance to 5-fluorouracil (5-FU) (50–1600 µM) and cisplatin (25–800 µM), reduced MMP, and caused the cell cycle arrest at G(0)/G(1) phase in a concentration-dependent manner. Hypoxia also concentration-dependently (5 % O(2), 3 % O(2), 1 % O(2)) reduced mRNA expression level of P-glycoprotein (P-gp), multidrug resistance protein (MRP1), lung resistance protein (LRP), and decreased the protein expression level of hypoxia-inducible factor-1α (HIF-1α), P-gp MRP1, and LRP. Following pretreatment with As(2)O(3) at a non-cytotoxic concentration re-sensitized the hypoxia (1 % O(2))-induced chemo-resistance to 5-FU and cisplatin in HBx-HepG2 cells. As(2)O(3) pretreatment also prevented MMP reduction and G(0)/G(1) arrest induced by hypoxia. Meanwhile, As(2)O(3) antagonized increase of HIF-1α protein induced by hypoxia, and it also suppresses the increase in expression levels of P-gp, MRP1, and LRP mRNA and proteins. In addition, As(2)O(3) in combination with 5-FU treatment caused up-regulation of DR5, caspase 3, caspase 8, and caspase 9, and down-regulation of BCL-2, but had no effect of DR4. CONCLUSIONS: Our results may suggest that As(2)O(3) re-sensitizes hypoxia-induced chemo-resistance in HBx-HepG2 via complex pathways, and As(2)O(3) may be a potential agent that given in combination with other anti-drugs for the treatment of HBV related HCC, which is resistant to chemotherapy. BioMed Central 2015-12-12 /pmc/articles/PMC4676851/ /pubmed/26692822 http://dx.doi.org/10.1186/s12935-015-0269-y Text en © Yu et al. 2015 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Primary Research Yu, Guifang Chen, Xuezhu Chen, Shudi Ye, Weipeng Hou, Kailian Liang, Min Arsenic trioxide reduces chemo-resistance to 5-fluorouracil and cisplatin in HBx-HepG2 cells via complex mechanisms |
title | Arsenic trioxide reduces chemo-resistance to 5-fluorouracil and cisplatin in HBx-HepG2 cells via complex mechanisms |
title_full | Arsenic trioxide reduces chemo-resistance to 5-fluorouracil and cisplatin in HBx-HepG2 cells via complex mechanisms |
title_fullStr | Arsenic trioxide reduces chemo-resistance to 5-fluorouracil and cisplatin in HBx-HepG2 cells via complex mechanisms |
title_full_unstemmed | Arsenic trioxide reduces chemo-resistance to 5-fluorouracil and cisplatin in HBx-HepG2 cells via complex mechanisms |
title_short | Arsenic trioxide reduces chemo-resistance to 5-fluorouracil and cisplatin in HBx-HepG2 cells via complex mechanisms |
title_sort | arsenic trioxide reduces chemo-resistance to 5-fluorouracil and cisplatin in hbx-hepg2 cells via complex mechanisms |
topic | Primary Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4676851/ https://www.ncbi.nlm.nih.gov/pubmed/26692822 http://dx.doi.org/10.1186/s12935-015-0269-y |
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