Hypoxia inducible factor-1 mediates upregulation of urokinase-type plasminogen activator receptor gene transcription during hypoxia in cervical cancer cells

Nishi,Hirotaka; Sasaki,Toru; Nagamitsu,Yuzo; Terauchi,Fumitoshi; Nagai,Takeshi; Nagao,Toshitaka; Isaka,Keiichi

doi:10.3892/or.2015.4449

Hypoxia inducible factor-1 mediates upregulation of urokinase-type plasminogen activator receptor gene transcription during hypoxia in cervical cancer cells

Authors:
- Hirotaka Nishi
- Toru Sasaki
- Yuzo Nagamitsu
- Fumitoshi Terauchi
- Takeshi Nagai
- Toshitaka Nagao
- Keiichi Isaka
View Affiliations

Affiliations: Department of Obstetrics and Gynecology, Tokyo Medical University, Shinjuku-ku, Tokyo 160-0023, Japan, Department of Anatomic Pathology, Tokyo Medical University, Shinjuku-ku, Tokyo 160-0023, Japan
Published online on: November 25, 2015 https://doi.org/10.3892/or.2015.4449
Pages: 992-998

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 occurs during development of cervical cancer and is considered to correlate with its invasion. Hypoxia mediates tumor cells to have more invasive property in a variety of cancers. Urokinase plasminogen activator receptor (uPAR) which mediates invasion is considered to be induced by hypoxia. We sought to determine the regulators of uPAR expression during hypoxia in cervical cancer. We showed that cervical cancer cell lines, CaSki and CA, were more invasive under hypoxic condition (1% O2) than under normoxic condition (20% O2) by invasion assays. Using western blot analysis, hypoxia enhanced the endogenous hypoxia-inducible factor (HIF)-1α and uPAR protein expression. uPAR mRNA level was also upregulated by hypoxia using real-time RT-PCR. Overexpression of HIF-1α which is induced by hypoxia activated the transcriptional activity of the uPAR promoter by luciferase assays. HIF-1 protein bound the putative HIF-1 response element on the uPAR promoter using electrophoretic mobility shift analysis, and additional luciferase assays show that this is essential for uPAR transactivation by HIF-1. HIF-1 overexpression enhanced the endogenous uPAR expression and introduction of siRNA for HIF-1α diminishes uPAR expression during hypoxia. These results indicate the upregulation of uPAR by hypoxia in cervical cancer cells is mediated through HIF-1. In cervical cancer tissues, we also demonstrated that uPAR protein expression was detected in cervical cancer but not in normal cervix or cervical intraepithelial neoplasia (CIN) by immunohistopathological staining. Our results provide evidence that regulation of uPAR expression by HIF-1 represents a mechanism for cervical cancer invasion during hypoxia.

View Figures

View References

1	Brizel DM, Scully SP, Harrelson JM, Layfield LJ, Bean JM, Prosnitz LR and Dewhirst MW: Tumor oxygenation predicts for the likelihood of distant metastases in human soft tissue sarcoma. Cancer Res. 56:941–943. 1996.PubMed/NCBI
2	Brizel DM, Sibley GS, Prosnitz LR, Scher RL and Dewhirst MW: Tumor hypoxia adversely affects the prognosis of carcinoma of the head and neck. Int J Radiat Oncol Biol Phys. 38:285–289. 1997. View Article : Google Scholar : PubMed/NCBI
3	Vaupel P, Thews O, Mayer A, Höckel S and Höckel M: Oxygenation status of gynecologic tumors: What is the optimal hemoglobin level? Strahlenther Onkol. 178:727–731. 2002. View Article : Google Scholar : PubMed/NCBI
4	Duong HS, Le AD, Zhang Q and Messadi DV: A novel 3-dimensional culture system as an in vitro model for studying oral cancer cell invasion. Int J Exp Pathol. 86:365–374. 2005. View Article : Google Scholar : PubMed/NCBI
5	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
6	Maity A and Solomon D: Both increased stability and transcription contribute to the induction of the urokinase plasminogen activator receptor (uPAR) message by hypoxia. Exp Cell Res. 255:250–257. 2000. View Article : Google Scholar : PubMed/NCBI
7	Montuori N, Mattiello A, Mancini A, Santoli M, Taglialatela P, Caputi M, Rossi G and Ragno P: Urokinase-type plasminogen activator up-regulates the expression of its cellular receptor through a post-transcriptional mechanism. FEBS Lett. 508:379–384. 2001. View Article : Google Scholar : PubMed/NCBI
8	Giatromanolaki A, Koukourakis MI, Sivridis E, Turley H, Talks K, Pezzella F, Gatter KC and Harris AL: Relation of hypoxia inducible factor 1 alpha and 2 alpha in operable non-small cell lung cancer to angiogenic/molecular profile of tumours and survival. Br J Cancer. 85:881–890. 2001. View Article : Google Scholar : PubMed/NCBI
9	Semenza GL: Hypoxia-inducible factors in physiology and medicine. Cell. 148:399–408. 2012. View Article : Google Scholar : PubMed/NCBI
10	Fukuda R, Zhang H, Kim JW, Shimoda L, Dang CV and Semenza GL: HIF-1 regulates cytochrome oxidase subunits to optimize efficiency of respiration in hypoxic cells. Cell. 129:111–122. 2007. View Article : Google Scholar : PubMed/NCBI
11	Nishi H, Nakada T, Kyo S, Inoue M, Shay JW and Isaka K: Hypoxia-inducible factor 1 mediates upregulation of telomerase (hTERT). Mol Cell Biol. 24:6076–6083. 2004. View Article : Google Scholar : PubMed/NCBI
12	McNally SJ, Harrison EM, Ross JA, Garden OJ and Wigmore SJ: Curcumin induces heme oxygenase 1 through generation of reactive oxygen species, p38 activation and phosphatase inhibition. Int J Mol Med. 19:165–172. 2007.
13	Kim J, Yu W, Kovalski K and Ossowski L: Requirement for specific proteases in cancer cell intravasation as revealed by a novel semiquantitative PCR-based assay. Cell. 94:353–362. 1998. View Article : Google Scholar : PubMed/NCBI
14	Yamano M, Fujii H, Takagaki T, Kadowaki N, Watanabe H and Shirai T: Genetic progression and divergence in pancreatic carcinoma. Am J Pathol. 156:2123–2133. 2000. View Article : Google Scholar : PubMed/NCBI
15	Lal A, Peters H, St Croix B, Haroon ZA, Dewhirst MW, Strausberg RL, Kaanders JH, van der Kogel AJ and Riggins GJ: Transcriptional response to hypoxia in human tumors. J Natl Cancer Inst. 93:1337–1343. 2001. View Article : Google Scholar : PubMed/NCBI
16	Koong AC, Denko NC, Hudson KM, Schindler C, Swiersz L, Koch C, Evans S, Ibrahim H, Le QT, Terris DJ, et al: Candidate genes for the hypoxic tumor phenotype. Cancer Res. 60:883–887. 2000.PubMed/NCBI
17	Graham CH, Forsdike J, Fitzgerald CJ and Macdonald-Goodfellow S: Hypoxia-mediated stimulation of carcinoma cell invasiveness via upregulation of urokinase receptor expression. Int J Cancer. 80:617–623. 1999. View Article : Google Scholar : PubMed/NCBI
18	Mazar AP, Henkin J and Goldfarb RH: The urokinase plasminogen activator system in cancer: Implications for tumor angiogenesis and metastasis. Angiogenesis. 3:15–32. 1999. View Article : Google Scholar
19	Bauer TW, Liu W, Fan F, Camp ER, Yang A, Somcio RJ, Bucana CD, Callahan J, Parry GC, Evans DB, et al: Targeting of urokinase plasminogen activator receptor in human pancreatic carcinoma cells inhibits c-Met- and insulin-like growth factor-I receptor-mediated migration and invasion and orthotopic tumor growth in mice. Cancer Res. 65:7775–7781. 2005.PubMed/NCBI
20	Reuning U, Sperl S, Kopitz C, Kessler H, Krüger A, Schmitt M and Magdolen V: Urokinase-type plasminogen activator (uPA) and its receptor (uPAR): Development of antagonists of uPA/uPAR interaction and their effects in vitro and in vivo. Curr Pharm Des. 9:1529–1543. 2003. View Article : Google Scholar : PubMed/NCBI
21	Degryse B, Sier CF, Resnati M, Conese M and Blasi F: PAI-1 inhibits urokinase-induced chemotaxis by internalizing the urokinase receptor. FEBS Lett. 505:249–254. 2001. View Article : Google Scholar : PubMed/NCBI
22	Ramont L, Pasco S, Hornebeck W, Maquart FX and Monboisse JC: Transforming growth factor-beta1 inhibits tumor growth in a mouse melanoma model by down-regulating the plasminogen activation system. Exp Cell Res. 291:1–10. 2003. View Article : Google Scholar : PubMed/NCBI
23	Fox SB, Taylor M, Grøndahl-Hansen J, Kakolyris S, Gatter KC and Harris AL: Plasminogen activator inhibitor-1 as a measure of vascular remodelling in breast cancer. J Pathol. 195:236–243. 2001. View Article : Google Scholar : PubMed/NCBI
24	Büchler P, Reber HA, Tomlinson JS, Hankinson O, Kallifatidis G, Friess H, Herr I and Hines OJ: Transcriptional regulation of urokinase-type plasminogen activator receptor by hypoxia-inducible factor 1 is crucial for invasion of pancreatic and liver cancer. Neoplasia. 11:196–206. 2009. View Article : Google Scholar : PubMed/NCBI
25	Montagne M, Naud JF, McDuff FO and Lavigne P: Toward the elucidation of the structural determinants responsible for the molecular recognition between Mad1 and Max. Biochemistry. 44:12860–12869. 2005. View Article : Google Scholar : PubMed/NCBI
26	Hu J, Banerjee A and Goss DJ: Assembly of b/HLH/z proteins c-Myc, Max, and Mad1 with cognate DNA: Importance of protein-protein and protein-DNA interactions. Biochemistry. 44:11855–11863. 2005. View Article : Google Scholar : PubMed/NCBI
27	Semov A, Marcotte R, Semova N, Ye X and Wang E: Microarray analysis of E-box binding-related gene expression in young and replicatively senescent human fibroblasts. Anal Biochem. 302:38–51. 2002. View Article : Google Scholar : PubMed/NCBI
28	Vervoorts J and Lüscher B: DNA binding of Myc/Max/Mad network complexes to oligonucleotides containing two E box elements: c-Myc/Max heterodimers do not bind DNA cooperatively. Biol Chem. 380:1121–1126. 1999. View Article : Google Scholar : PubMed/NCBI
29	Koshiji M, Kageyama Y, Pete EA, Horikawa I, Barrett JC and Huang LE: HIF-1alpha induces cell cycle arrest by functionally counteracting Myc. EMBO J. 23:1949–1956. 2004. View Article : Google Scholar : PubMed/NCBI
30	Powis G and Kirkpatrick L: Hypoxia inducible factor-1alpha as a cancer drug target. Mol Cancer Ther. 3:647–654. 2004.PubMed/NCBI