Dominant-Negative Hypoxia-Inducible Factor-1α Reduces Tumorigenicity of Pancreatic Cancer Cells through the Suppression of Glucose Metabolism

Chen, Jian; Zhao, Shen; Nakada, Kunihiro; Kuge, Yuji; Tamaki, Nagara; Okada, Futoshi; Wang, Jingxin; Shindo, Masanobu; Higashino, Fumihiro; Takeda, Kohji; Asaka, Masahiro; Katoh, Hiroyuki; Sugiyama, T.; Hosokawa, Masuo; Kobayashi, Masanobu

doi:10.1016/s0002-9440(10)63924-7

Cited by 169 publications

(126 citation statements)

References 34 publications

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“…Enhanced glycolytic metabolism through increased glucose uptake via GLUT-1 has been shown to prevent cell damage during hypoxia (Chen et al, 2003). In addition, overexpression of GLUT-1 prevented hypoxia-induced apoptosis also independent of extracellular glucose concentrations by inhibiting stress-activated protein kinase activation (Lin et al, 2000).…”

Section: N H H-gd A204 A673mentioning

confidence: 99%

Role of hypoxia inducible factor-1 alpha in modulation of apoptosis resistance

et al. 2006

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Hypoxia inducible factor-1 (HIF-1) is the major transcription factor and key regulator of adoptive responses to hypoxia. Although it usually promotes tumor cell survival under hypoxia, it has also been implied to trigger apoptosis. Although the impact of hypoxia has been extensively studied in many adult solid tumors, its role in most childhood tumors, for example, in rhabdomyosarcoma (RMS) or Ewing sarcoma (ES), has not yet been addressed. Here, we report that hypoxia protects A204 RMS and A673 ES cells against anticancer drug-or tumor necrosis factor-related apoptosis-inducing ligandinduced apoptosis and that Hif-1a plays a key role in conferring apoptosis resistance under hypoxia. Although a functional HIF-1 pathway and proapoptotic proteins such as p53 and Bcl-2/E1B 19 kDa interacting protein 3 were activated under hypoxia in both A204 RMS and A673 ES cells, these cells remained refractory to apoptosis. Concomitant analysis of antiapoptotic proteins revealed that hypoxia induced expression of Bcl-2 and inhibitor of apoptosis proteins (IAP)-2 as well as proteins associated with anaerobic metabolism such as the glucose transporter protein GLUT-1 and the glycolytic enzyme Aldolase A. Specific downregulation of Hif-1a by RNA interference significantly enhanced apoptosis under hypoxia by preventing the hypoxia-mediated increase in GLUT-1 expression without altering expression levels of the antiapoptotic proteins Bcl-2 or cIAP-2. Moreover, glucose deprivation-induced apoptosis of A204 RMS and A673 ES cells was inhibited under hypoxic conditions in a Hif-1a-dependent manner. As GLUT-1 was induced via Hif-1a under hypoxia in A204 RMS and A673 ES, these findings suggest that the Hif-1a-mediated increase in glucose uptake plays an important role in conferring apoptosis resistance. Thus, hypoxia-inducible genes may represent novel targets for therapeutic intervention in some pediatric tumors, which warrants further investigation.

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Section: N H H-gd A204 A673mentioning

confidence: 99%

Role of hypoxia inducible factor-1 alpha in modulation of apoptosis resistance

et al. 2006

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“…35 In this respect, it is worth noting that HIF-1a can mediate crosstalk between hypoxia and glucose metabolism via glucose response elements. 36,37 Since a significant proportion of transfected HUVECs did not manifest evidence of apoptotic changes by TUNEL assay and activated caspase-3 immunostaining after A/R in the current study, it is probable that other antiapoptotic proteins distinct from HIF-1a may also have conferred some cytoprotection against anoxic stress, as has been shown with respect to CD105 38 and Mcl-1. 39 In summary, we have employed RNAi targeting the HIF-1a gene in primary cultured HUVECs and have demonstrated the role of HIF-1a as an antiapoptotic protein in a model of anoxic stress.…”

Section: Discussionmentioning

confidence: 50%

Antiapoptotic action of hypoxia-inducible factor-1α in human endothelial cells

Liu

et al. 2004

Laboratory Investigation

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Hypoxia-inducible factor-1 (HIF-1) is the major transcription factor involved in the adaptive response to hypoxia and consists of HIF-1a and HIF-1b subunits. Indirect evidence suggests that HIF-1a may exert both proapoptotic and antiapoptotic actions in response to hypoxia. In this study, we evaluated the effects of RNA interference (RNAi) targeting HIF-1a messenger RNA (mRNA) on apoptosis in primary cultured human umbilical vascular endothelial cells (HUVECs) exposed to anoxia and reoxygenation (A/R). HUVECs were transfected with specific 21-nt small interfering RNA (siRNA) duplexes targeting HIF-1a mRNA sequences or scrambled RNA duplexes and subjected either to normoxia for 251/2 h or to anoxia for 11/2 h, and subsequently normoxia for 24 h (A/R). Control samples were subjected to A/R but not transfected. HUVECs apoptosis was evaluated by Tdt-mediated dUTP nick end-labeling (TUNEL) assay and by activated caspase-3 immunostaining and immunoblotting. The efficacy of RNAi was assessed by knockdown of HIF-1a mRNA and protein expression via in situ hybridization, real-time quantitative PCR, immunohistochemistry, and Western blotting. When compared with normoxic cultures, A/R significantly upregulated HIF-1a mRNA and protein expression in HUVECs, but did not appreciably alter the percentage of apoptotic cells. In contrast, a significantly greater proportion of HUVECs transfected with specific siRNA duplexes and exposed to A/R demonstrated evidence of apoptosis when compared with nontransfected cells. Transfection with specific siRNA duplexes knocked down HIF-1a mRNA and protein expression in A/R-treated cells by approximately 60%, whereas transfection with scrambled siRNA duplexes had no noticeable effect on HIF-1a expression. These findings strongly suggest that HIF-1a exerts an antiapoptotic role in HUVECs stressed by anoxia.

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“…In favor of this hypothesis are results from a recently published study demonstrating that KRAS activation promotes KLF5 up-regulation in human colon cancer (HCT116) cells (13). Moreover, because hypoxia-mediated and cytokine-mediated activation of the oncogenic transcription factor hypoxia-inducible factor-1α (HIF-1α) is frequently encountered in pancreatic tumors, we also hypothesized that the interleukin (IL)-1β system and/or HIF-1α may be involved in the regulation of KLF5 expression (14)(15)(16)(17)(18). We therefore sought to further define the expression and regulation of KLF5 in human pancreatic cancer cells to expand our knowledge on this particular transcription factor and to potentially reveal a novel molecular therapeutic target.…”

Section: Introductionmentioning

confidence: 95%

Up-Regulation of Krüppel-Like Factor 5 in Pancreatic Cancer Is Promoted by Interleukin-1β Signaling and Hypoxia-Inducible Factor-1α

Mori

Moser

Lang

et al. 2009

Molecular Cancer Research

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Krüppel-like factor 5 (KLF5) is a transcription factor involved in cell transformation, proliferation, and carcinogenesis that can be up-regulated by RAS mutations. However, controversy persists as to whether it functions as a tumor suppressor or as an oncogene. Because KRAS is frequently mutated in pancreatic cancer, we investigated the regulation of KLF5 in this cancer entity. Our results show that KLF5 is overexpressed in pancreatic cancer cells and exceeds KLF5 expression of KRAS-mutated colon cancer cells. Surprisingly, inhibition of B-Raf/C-Raf or MAPK/Erk did not reduce KLF5 levels, suggesting that KLF5 expression is not promoted by KRAS-Raf-MEK-Erk signaling in pancreatic cancer. This finding is in striking contrast to reports on MEK-Erk-mediated KLF5 induction in colon cancer cells. Moreover, KLF5 expression levels neither correlated with the mutational status of KRAS nor with MEK phosphorylation in pancreatic cancer cells. Importantly, KLF5 was significantly up-regulated by interleukin (IL)-1β or hypoxia. The IL-1 β-mediated induction of KLF5 was diminished by blocking the p38 pathway. In addition, blocking IL-1R reduced the constitutive KLF5 expression, suggesting an autocrine activation loop. Moreover, KLF5 coimmunoprecipitated with hypoxia-inducible factor-1α (HIF-1α) and HIF-1α siRNA reduced constitutive KLF5. Similarly, KLF5 siRNA reduced the expression of the HIF-1α target gene GLUT-1. Furthermore, KLF5 expression was significantly elevated by high cell density, by anchorage-independent cell growth, and in tumor spheroids. Down-regulation of KLF5 by RNAi reduced the expression of the target genes, survivin, and platelet-derived growth factor-A. In conclusion, overexpression of KLF5 in human pancreatic cancer cells is not mediated by KRAS/Raf/ MAPK/Erk signaling, but involves the IL-1β/IL-1R system, p38, and the transcription factor

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Dominant-Negative Hypoxia-Inducible Factor-1α Reduces Tumorigenicity of Pancreatic Cancer Cells through the Suppression of Glucose Metabolism

Cited by 169 publications

References 34 publications

Role of hypoxia inducible factor-1 alpha in modulation of apoptosis resistance

Role of hypoxia inducible factor-1 alpha in modulation of apoptosis resistance

Antiapoptotic action of hypoxia-inducible factor-1α in human endothelial cells

Up-Regulation of Krüppel-Like Factor 5 in Pancreatic Cancer Is Promoted by Interleukin-1β Signaling and Hypoxia-Inducible Factor-1α

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