HAF drives the switch of HIF-1α to HIF-2α by activating the NF-κB pathway, leading to malignant behavior of T24 bladder cancer cells

Guan,Zhenfeng; Ding,Chen; Du,Yiqing; Zhang,Kai; Zhu,Jian Ning; Zhang,Tingting; He,Dalin; Xu,Shan; Wang,Xinyang; Fan,Jinhai

doi:10.3892/ijo.2013.2210

HAF drives the switch of HIF-1α to HIF-2α by activating the NF-κB pathway, leading to malignant behavior of T24 bladder cancer cells

Authors:
- Zhenfeng Guan
- Chen Ding
- Yiqing Du
- Kai Zhang
- Jian Ning Zhu
- Tingting Zhang
- Dalin He
- Shan Xu
- Xinyang Wang
- Jinhai Fan
View Affiliations

Affiliations: Oncology Research Lab, Key Laboratory of Environment and Genes Related to Diseases, Ministry of Education, Xi'an, P.R. China, Department of Urology, The First Affiliated Hospital of Medical College of Xi'an Jiaotong University, Xi'an, P.R. China
Published online on: December 6, 2013 https://doi.org/10.3892/ijo.2013.2210
Pages: 393-402
Copyright: © Guan et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Hypoxia is a characteristic feature of solid tumors, leading to malignant behavior. During this process, HIF family members (HIFs) and the NF-κB pathway are activated. In addition, the hypoxia-associated factor (HAF) is reported to participate in the regulation of HIFs. However, the precise relationship among HIFs, HAF and the NF-κB pathway in bladder cancer (BC) remains unknown. In the current investigation, T24 BC cells were exposed to hypoxia, or by plasmid transfection to overexpress HAF or RelA (P65) to demonstrate their roles. The results indicate that hypoxia leads to the elevation of HAF plus activation of the NF-κB pathway, accompanied by the switch of HIF-1α to HIF-2α, resulting in the enhanced ability of malignancy in T24 cells. In order to further demonstrate the significance of this switch, HIF-1α and HIF-2α were co-transfected into T24 cells with HIF-β, respectively. The following results indicate that the T24hif-2α/β cells show enhanced ability of malignancy, accompanied by the maintenance of stem-cell markers, but the T24hif-1α/β cells show higher expression of metabolism-related genes. Boyden assays and wound-healing assays indicate the enhanced ability of malignancy for T24hif-2α/β. Thus, we conclude that on the hypoxic microenvironment, the switching of HIF-1α to HIF-2α, which is driven by HAF through activating the NF-κB pathway, contributes to the malignancy of T24 cells, accompanied by the maintenance of stem-cell markers. This provides us an avenue for understanding the progression of bladder cancer.

View Figures

View References

1.	Nathke I and Rocha S: Antagonistic crosstalk between APC and HIF-1alpha. Cell Cycle. 10:1545–1547. 2011. View Article : Google Scholar : PubMed/NCBI
2.	Yoo YG, Christensen J, Gu J and Huang LE: HIF-1alpha mediates tumor hypoxia to confer a perpetual mesenchymal phenotype for malignant progression. Sci Signal. 4:pt42011.PubMed/NCBI
3.	Haase VH: Hypoxia-inducible factors in the kidney. Am J Physiol Renal Physiol. 291:F271–F281. 2006. View Article : Google Scholar : PubMed/NCBI
4.	Henze AT and Acker T: Feedback regulators of hypoxia-inducible factors and their role in cancer biology. Cell Cycle. 9:2749–2763. 2010. View Article : Google Scholar : PubMed/NCBI
5.	Berra E, Ginouves A and Pouyssegur J: The hypoxia-inducible-factor hydroxylases bring fresh air into hypoxia signalling. EMBO Rep. 7:41–45. 2006. View Article : Google Scholar : PubMed/NCBI
6.	Koh MY and Powis G: Passing the baton: the HIF switch. Trends Biochem Sci. 37:364–372. 2012. View Article : Google Scholar : PubMed/NCBI
7.	Yoo YG, Christensen J and Huang LE: HIF-1alpha confers aggressive malignant traits on human tumor cells independent of its canonical transcriptional function. Cancer Res. 71:1244–1252. 2011. View Article : Google Scholar : PubMed/NCBI
8.	Loboda A, Jozkowicz A and Dulak J: HIF-1 and HIF-2 transcription factors - similar but not identical. Mol Cells. 29:435–442. 2010. View Article : Google Scholar : PubMed/NCBI
9.	Majmundar AJ, Wong WJ and Simon MC: Hypoxia-inducible factors and the response to hypoxic stress. Mol Cell. 40:294–309. 2010. View Article : Google Scholar : PubMed/NCBI
10.	Rohwer N and Cramer T: Hypoxia-mediated drug resistance: novel insights on the functional interaction of HIFs and cell death pathways. Drug Resist Updat. 14:191–201. 2011. View Article : Google Scholar : PubMed/NCBI
11.	Heddleston JM, Li Z, Lathia JD, Bao S, Hjelmeland AB and Rich JN: Hypoxia inducible factors in cancer stem cells. Br J Cancer. 102:789–795. 2010. View Article : Google Scholar : PubMed/NCBI
12.	Tafani M, Schito L, Pellegrini L, et al: Hypoxia-increased RAGE and P2X7R expression regulates tumor cell invasion through phosphorylation of Erk1/2 and Akt and nuclear translocation of NF-{kappa}B. Carcinogenesis. 32:1167–1175. 2011.PubMed/NCBI
13.	Yamakuchi M, Yagi S, Ito T and Lowenstein CJ: MicroRNA-22 regulates hypoxia signaling in colon cancer cells. PLoS One. 6:e202912011. View Article : Google Scholar : PubMed/NCBI
14.	Zhdanov AV, Dmitriev RI, Golubeva AV, Gavrilova SA and Papkovsky DB: Chronic hypoxia leads to a glycolytic phenotype and suppressed HIF-2 signaling in PC12 cells. Biochim Biophys Acta. 1830:3553–3569. 2013. View Article : Google Scholar : PubMed/NCBI
15.	Petrella BL, Lohi J and Brinckerhoff CE: Identification of membrane type-1 matrix metalloproteinase as a target of hypoxia-inducible factor-2 alpha in von Hippel-Lindau renal cell carcinoma. Oncogene. 24:1043–1052. 2005. View Article : Google Scholar : PubMed/NCBI
16.	Koh MY, Lemos RJ, Liu X and Powis G: The hypoxia-associated factor switches cells from HIF-1alpha- to HIF-2alpha-dependent signaling promoting stem cell characteristics, aggressive tumor growth and invasion. Cancer Res. 71:4015–4027. 2011. View Article : Google Scholar
17.	Li Z and Rich JN: Hypoxia and hypoxia inducible factors in cancer stem cell maintenance. Curr Top Microbiol Immunol. 345:21–30. 2010.PubMed/NCBI
18.	Yeung TM, Gandhi SC and Bodmer WF: Hypoxia and lineage specification of cell line-derived colorectal cancer stem cells. Proc Natl Acad Sci USA. 108:4382–4387. 2011. View Article : Google Scholar : PubMed/NCBI
19.	Yeramian A, Santacana M, Sorolla A, et al: Nuclear factor-kappaB2/p100 promotes endometrial carcinoma cell survival under hypoxia in a HIF-1alpha independent manner. Lab Invest. 91:859–871. 2011. View Article : Google Scholar : PubMed/NCBI
20.	Xue Y, Li NL, Yang JY, Chen Y, Yang LL and Liu WC: Phosphatidylinositol 3’-kinase signaling pathway is essential for Rac1-induced hypoxia-inducible factor-1(alpha) and vascular endothelial growth factor expression. Am J Physiol Heart Circ Physiol. 300:H2169–H2176. 2011.
21.	Nys K, Maes H, Dudek AM and Agostinis P: Uncovering the role of hypoxia inducible factor-1alpha in skin carcinogenesis. Biochim Biophys Acta. 1816:1–12. 2011.PubMed/NCBI
22.	Ghosh G, Wang VY, Huang DB and Fusco A: NF-kappaB regulation: lessons from structures. Immunol Rev. 246:36–58. 2012. View Article : Google Scholar : PubMed/NCBI
23.	van Uden P, Kenneth NS, Webster R, Muller HA, Mudie S and Rocha S: Evolutionary conserved regulation of HIF-1beta by NF-kappaB. PLoS Genet. 7:e10012852011.PubMed/NCBI
24.	Sun SC: The noncanonical NF-kappaB pathway. Immunol Rev. 246:125–140. 2012.
25.	Paliwal P, Arora D and Mishra AK: Epithelial mesenchymal transition in urothelial carcinoma: twist in the tale. Indian J Pathol Microbiol. 55:443–449. 2012. View Article : Google Scholar : PubMed/NCBI
26.	Jing Y, Han Z, Zhang S, Liu Y and Wei L: Epithelial-mesenchymal transition in tumor microenvironment. Cell Biosci. 1:292011. View Article : Google Scholar : PubMed/NCBI
27.	Wu ST, Sun GH, Hsu CY, et al: Tumor necrosis factor-alpha induces epithelial-mesenchymal transition of renal cell carcinoma cells via a nuclear factor kappa B-independent mechanism. Exp Biol Med. 236:1022–1029. 2011. View Article : Google Scholar
28.	Harhaj EW and Dixit VM: Regulation of NF-kappaB by deubiquitinases. Immunol Rev. 246:107–124. 2012.PubMed/NCBI
29.	Gilmore TD and Wolenski FS: NF-kappaB: where did it come from and why? Immunol Rev. 246:14–35. 2012. View Article : Google Scholar : PubMed/NCBI
30.	Kanarek N and Ben-Neriah Y: Regulation of NF-kappaB by ubiquitination and degradation of the IkappaBs. Immunol Rev. 246:77–94. 2012. View Article : Google Scholar : PubMed/NCBI
31.	Brennan P and O’Neill LA: 2-mercaptoethanol restores the ability of nuclear factor kappa B (NF kappa B) to bind DNA in nuclear extracts from interleukin 1-treated cells incubated with pyrollidine dithiocarbamate (PDTC). Evidence for oxidation of glutathione in the mechanism of inhibition of NF kappa B by PDTC. Biochem J. 320:975–981. 1996.
32.	Hayakawa M, Miyashita H, Sakamoto I, et al: Evidence that reactive oxygen species do not mediate NF-kappaB activation. EMBO J. 22:3356–3366. 2003. View Article : Google Scholar : PubMed/NCBI
33.	Julien S, Puig I, Caretti E, et al: Activation of NF-kappaB by Akt upregulates Snail expression and induces epithelium mesenchyme transition. Oncogene. 26:7445–7456. 2007. View Article : Google Scholar : PubMed/NCBI
34.	Vaupel P and Mayer A: Hypoxia in cancer: significance and impact on clinical outcome. Cancer Metastasis Rev. 26:225–239. 2007. View Article : Google Scholar : PubMed/NCBI
35.	Weidemann A and Johnson RS: Biology of HIF-1alpha. Cell Death Differ. 15:621–627. 2008. View Article : Google Scholar : PubMed/NCBI
36.	Saito T and Kawaguchi H: HIF-2alpha as a possible therapeutic target of osteoarthritis. Osteoarthritis Cartilage. 18:1552–1556. 2010. View Article : Google Scholar : PubMed/NCBI
37.	Uchida T, Rossignol F, Matthay MA, et al: Prolonged hypoxia differentially regulates hypoxia-inducible factor (HIF)-1alpha and HIF-2alpha expression in lung epithelial cells: implication of natural antisense HIF-1alpha. J Biol Chem. 279:14871–14878. 2004. View Article : Google Scholar
38.	Myllyharju J and Schipani E: Extracellular matrix genes as hypoxia-inducible targets. Cell Tissue Res. 339:19–29. 2010. View Article : Google Scholar : PubMed/NCBI
39.	Jiang H, Zhu Y, Xu H, Sun Y and Li Q: Activation of hypoxiainducible factor-1alpha via nuclear factor-kappa B in rats with chronic obstructive pulmonary disease. Acta Biochim Biophys Sin. 42:483–488. 2010. View Article : Google Scholar : PubMed/NCBI
40.	Levidou G, Saetta AA, Korkolopoulou P, et al: Clinical significance of nuclear factor (NF)-kappaB levels in urothelial carcinoma of the urinary bladder. Virchows Arch. 452:295–304. 2008. View Article : Google Scholar : PubMed/NCBI
41.	Gorlach A and Bonello S: The cross-talk between NF-kappaB and HIF-1: further evidence for a significant liaison. Biochem J. 412:e17–e19. 2008.PubMed/NCBI
42.	Taylor CT and Cummins EP: The role of NF-kappaB in hypoxia-induced gene expression. Ann NY Acad Sci. 1177:178–184. 2009. View Article : Google Scholar : PubMed/NCBI
43.	van Uden P, Kenneth NS and Rocha S: Regulation of hypoxiainducible factor-1alpha by NF-kappaB. Biochem J. 412:477–484. 2008.PubMed/NCBI
44.	Nam SY, Ko YS, Jung J, et al: A hypoxia-dependent upregulation of hypoxia-inducible factor-1 by nuclear factor-kappaB promotes gastric tumour growth and angiogenesis. Br J Cancer. 104:166–174. 2011. View Article : Google Scholar : PubMed/NCBI