Molecular characterizations of derivatives of HCT116 colorectal cancer cells that are resistant to the chemotherapeutic agent 5-fluorouracil

Angelis, Paula M. De; Fjell, Bjørg; Kravik, Katherine; Haug, Terje; Tunheim, Siv H.; Reichelt, Wenche H.; Beigi, Marzieh; Clausen, O. P. F.; Galteland, Eivind; Stokke, Trond

doi:10.3892/ijo.24.5.1279

Cited by 65 publications

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“…BNIP3 is overexpressed in human tumors and human tumor cell lines (11,21,40,41) and in primary neonatal rat cardiac myocytes exposed to hypoxia (14,22). At oxygen tensions used in cell culture experiments (which may or may not reflect the conditions under which these cells would exist in vivo), most tissues have undetectable levels of BNIP3 but activate transcription during hypoxia through a 5Ј promoter of HIF-1 binding site (49).…”

Section: Discussionmentioning

confidence: 99%

“…We have also shown that NO regulates the intestinal and hepatic expression of BNIP3 in a rat model in vivo (55). Despite these studies and their implication of BNIP3 in cell death, the ultimate significance of the regulation of the cell death inducer BNIP3 by NO still needs to be established.BNIP3 is overexpressed in human tumors and human tumor cell lines (11,21,40,41) and in primary neonatal rat cardiac myocytes exposed to hypoxia (14,22). At oxygen tensions used in cell culture experiments (which may or may not reflect the conditions under which these cells would exist in vivo), most tissues have undetectable levels of BNIP3 but activate transcription during hypoxia through a 5Ј promoter of HIF-1 binding site (49).…”

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confidence: 99%

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Expression and subcellular localization of BNIP3 in hypoxic hepatocytes and liver stress

Metukuri¹,

Beer‐Stolz²,

Namas³

et al. 2009

American Journal of Physiology-Gastrointestinal and Liver Physi

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We have previously demonstrated that the Bcl-2/adenovirus EIB 19-kDa interacting protein 3 (BNIP3), a cell death-related member of the Bcl-2 family, is upregulated in vitro and in vivo in both experimental and clinical settings of redox stress and that nitric oxide (NO) downregulates its expression. In this study we sought to examine the expression and localization of BNIP3 in murine hepatocytes and in a murine model of hemorrhagic shock (HS) and ischemia-reperfusion (I/R). Freshly isolated mouse hepatocytes were exposed to 1% hypoxia for 6 h followed by reoxygenation for 18 h, and protein was isolated for Western blot analysis. Hepatocytes grown on coverslips were fixed for localization studies. Similarly, livers from surgically cannulated C57Bl/6 mice and from mice cannulated and subjected to 1-4 h of HS were processed for protein isolation and Western blot analysis. In hepatocytes, BNIP3 was expressed constitutively but was upregulated under hypoxic conditions, and this upregulation was countered by treatment with a NO donor. Surprisingly, BNIP3 was localized in the nucleus of normoxic hepatocytes, in the cytoplasm following hypoxia, and again in the nucleus following reoxygenation. Upregulation of BNIP3 partially required p38 MAPK activation. BNIP3 contributed to hypoxic injury in hepatocytes, since this injury was diminished by knockdown of BNIP3 mRNA. Hepatic BNIP3 was also upregulated in two different models of liver stress in vivo, suggesting that a multitude of inflammatory stresses can lead to the modulation of BNIP3. In turn, the upregulation of BNIP3 appears to be one mechanism of hepatocyte cell death and liver damage in these settings. redox stress; nitric oxide; hypoxia; cell death; inflammation THE BCL-2/ADENOVIRUS EIB 19-kDa interacting protein 3 (BNIP3, formerly known as NIP3) was first identified in a yeast twohybrid screen as a protein capable of interacting with the adenovirus protein E1B 19 kDa, a homolog of Bcl-2 (4). BNIP3 is a membrane-associated protein localized to mitochondria and other cytoplasmic membrane structures and is widely expressed in a large number of mouse and human tissues (8). BNIP3 appears to have an overall resemblance to several BH3-containing Bcl-2 family proteins such as BIK, BID, HRK, and BAD, in which the BH3 domain plays an important role in the induction of apoptosis. However, BNIP3 has been implicated in both apoptotic and necrotic cell death (5). For instance, the expression of BNIP3 (both mRNA and protein) is induced and related to hypoxia-induced apoptosis in a number of human and animal cell lines, including CHO-K1 (Chinese hamster ovary), CV-1 (monkey kidney), Rat-1 (rat fibroblast), PAM212 (human epithelial), HepG2 (human hepatocellular carcinoma), and ECV-304 (human bladder carcinoma) (5). Moreover, BNIP3 and Nix [a BNIP3 homolog sharing both structural and functional similarity (7), also known as BNIP3L (30), BNIP3␣ (50), and B5 (35)] are the only members of the Bcl-2 family of apoptotic factors induced in response to hypoxia (5). On the other hand, BNI...

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Expression and subcellular localization of BNIP3 in hypoxic hepatocytes and liver stress

Metukuri¹,

Beer‐Stolz²,

Namas³

et al. 2009

American Journal of Physiology-Gastrointestinal and Liver Physi

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“…[73][74][75] Treatment with Aza-dC restores gene expression and the susceptibility to hypoxia-induced cell death. 73 BNIP3 is downregulated in an oxaliplatin-resistant colon cancer cell line compared with the sensitive cell line identified by microarray, 76 and in 5-FU-resistant colorectal cells, 77 suggesting that BNIP3 is an important regulator in the induction of drug-induced cell death in colon cancer. Some colon cancer cell lines have unmethylated bnip3 gene but still have no induction of BNIP3 expression under hypoxic conditions.…”

Section: Bnip3mentioning

confidence: 99%

The role of Bcl-2 family member BNIP3 in cell death and disease: NIPping at the heels of cell death

2009

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Bcl-2 nineteen-kilodalton interacting protein (BNIP3) is a BH-3-only Bcl-2 family member whose expression levels increase during stress such as hypoxia through hypoxia-inducing factor-1-dependent or -independent mechanisms. When BNIP3 expression is induced, it localizes to the mitochondria and triggers a loss of membrane potential, and an increase in the reactive oxygen species production, which often leads to cell death. Cells under normal growth conditions suppress BNIP3 expression through transcriptional repression. There is considerable debate in the literature regarding what type of cell death is induced by BNIP3. It has been observed that BNIP3 could induce necrosis, autophagy and/or apoptosis. In contrast, other studies indicate that BNIP3 could promote cell survival. Besides its cell death regulation, BNIP3 plays a key role in the pathogenicity of many diseases. In cardiac infarction, loss of BNIP3 expression has been shown to reduce the number of damaged cardiomyocytes after ischemia and reperfusion. BNIP3 expression also plays an important role in the deregulation of cell death in many cancers. In this review, we will discuss the different and often contradictory mechanisms of BNIP3 regulation of cell death and the role that BNIP3 may play in diseases. Bcl-2 nineteen-kilodalton interacting protein (BNIP3) belongs to the Bcl-2 homology domain (BH3)-only Bcl-2 family members because it only contains a putative BH3 domain. 1 The other major domains found in BNIP3 are the PEST domain that targets BNIP3 for degradation, a conserved domain that is conserved from Caenorhabditis elegans to humans and a transmembrane (TM) domain, which targets BNIP3 to the mitochondria (Figure 1). 2 BH3-only containing proteins act as rheostats regulating apoptosis through their BH3 domain by binding to antiapoptotic Bcl-2 family members. However, BNIP3 differs from these members, as its BH3 domain fails to interact with antiapoptotic Bcl-2 family members. 3 Furthermore, deletions of the BH3 domain fail to affect the ability of BNIP3 to induce cell death. 4 Unlike other BH3-only family members, BNIP3 interacts with Bcl-2 and Bcl-X L through its TM domain and N terminus (amino acids 1-49). 3 Deletion of the TM domain blocks the ability of BNIP3 to induce cell death. 4 BNIP3 migrates at 30 and 60 kDa, indicating that BNIP3 is a protein that forms homodimers. This homodimerization is primarily through the TM domain of BNIP3. The unique structure of the TM domain suggests that BNIP3 dimers could act as proton channels in the outer mitochondrial membrane increasing ion conductance. 5 Serine 172 and histidine (His) 173 are residues that are present in the dimerization interface of BNIP3. These residues interact by hydrogen bonds and are essential for dimer formation. 6 Furthermore, mutation of the His 173 to alanine completely abrogated the ability of BNIP3 to induce cell death. 7 Bcl-2 and Bcl-X L can compete for binding to the TM domain, which blocks BNIP3 homodimerization and abrogates the ability of BNIP3 to induce cell death (Fi...

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“…Therefore silencing of BNIP3 by methylation has been suggested to be essential for the survival of pancreatic cancer cells under hypoxic condition. Hypoxia-induced expression of BNIP3 is also impaired in other cancer cell lines, although there are differences in the extent of methylation and silencing (de Angelis et al, 2004;Kennedy et al, 2000). Murai and colleagues detected methylation of BNIP3 in 66% of primary colorectal and 49% of gastric cancers, 15% of acute lymphocytic and 17% of acute myelogenous leukaemias, and about 21% of multiple myelomas, but not in adjacent normal tissues (Murai et al, 2005a;2005b).…”

Section: Introductionmentioning

confidence: 99%

Silencing of BNIP3 Results from Promoter Methylation by DNA Methyltransferase 1 Induced by the Mitogen-Activated Protein Kinase Pathway

Lee

Paik

2011

Molecules and Cells

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We have previously shown that Ras mediates NO-induced BNIP3 expression via the MEK-ERK-HIF-1 pathway in mouse macrophages, and that NO-induced death results at least in part from the induction of BNIP3. In the present study, we describe another aspect of Ras regulation of BNIP3 expression in pancreatic cancer cells. Human BNIP3 promoter-driven luciferase activity was efficiently induced by activated Ras in AsPC-1, Miapaca-2, PK-1 and PANC-1 cells. However, expression of endogenous BNIP3 was not induced, and BNIP3 up-regulation by hypoxia was also inhibited. Treatment of the cells with the DNMT inhibitor, 5-aza-2-deoxycytidine, restored BNIP3 induction, indicating that DNA methylation of the BNIP3 promoter was responsible for the inhibition of BNIP3 induction. Furthermore, inhibition of the MEK pathway with U0126 reduced DNMT1 expression, but not that of DNMT3a and 3b, and restored the hypoxia-inducibility of BNIP3, suggesting that the DNA methylation of the BNIP3 promoter was mediated by DNMT1 via the MEK pathway.

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Molecular characterizations of derivatives of HCT116 colorectal cancer cells that are resistant to the chemotherapeutic agent 5-fluorouracil

Cited by 65 publications

References 0 publications

Expression and subcellular localization of BNIP3 in hypoxic hepatocytes and liver stress

Expression and subcellular localization of BNIP3 in hypoxic hepatocytes and liver stress

The role of Bcl-2 family member BNIP3 in cell death and disease: NIPping at the heels of cell death

Silencing of BNIP3 Results from Promoter Methylation by DNA Methyltransferase 1 Induced by the Mitogen-Activated Protein Kinase Pathway

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