CoCl2 increases the expression of hypoxic markers HIF‑1α, VEGF and CXCR4 in breast cancer MCF‑7 cells

Li,Qing; Ma,Rong; Zhang,Mei

doi:10.3892/ol.2017.7369

CoCl2 increases the expression of hypoxic markers HIF‑1α, VEGF and CXCR4 in breast cancer MCF‑7 cells

Authors:
- Qing Li
- Rong Ma
- Mei Zhang
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Affiliations: Department of Breast and Thyroid Surgery, Shandong Provincial Qianfoshan Hospital, Shandong University, Jinan, Shandong 250000, P.R. China
Published online on: November 8, 2017 https://doi.org/10.3892/ol.2017.7369
Pages: 1119-1124

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Abstract

The aim of the present study was to investigate the effect of a hypoxic environment on the biological behavior of breast cancer MCF‑7 cells, using CoCl2 to mimic the hypoxia model in breast cancer cells. Using 50, 100, 150 and 200 µM CoCl2 as a hypoxic inducer, a hypoxic model was established in MCF‑7 cells in vitro. MTT, reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR), terminal deoxynucleotidyl transferase dUTP nick‑end labeling (TUNEL) and western blotting assays were performed to detect MCF‑7 cell proliferation under hypoxic conditions and the expression of the hypoxic markers hypoxia‑inducible factor‑1α (HIF‑1α), vascular endothelial growth factor (VEGF) and C‑X‑C motif chemokine receptor 4 (CXCR4) mRNA and that of the associated proteins. The RT‑qPCR results revealed that there were no obvious changes in the expression of HIF‑1α mRNA; however, the expression of CXCR4 and VEGF mRNA increased significantly following treatment with different CoCl2 concentrations (P<0.05). The results of western blotting identified that CoCl2 significantly induced the expression of HIF‑1α, CXCR4 and VEGF proteins (P<0.05). The MTT assay revealed that different concentrations of CoCl2 inhibited the proliferation of MCF‑7 cells. The TUNEL assay demonstrated that CoCl2 was able to trigger apoptosis of MCF‑7 cells. Therefore, the results of the present study identified that CoCl2 is able to control MCF‑7 cell proliferation and apoptosis, also increasing the expression of HIF‑1α, CXCR4 and VEGF. The present study may aid the discovery of a novel method to prevent cell damage and decrease cell proliferation in order to prevent the occurrence and development of breast cancer.

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1	Hales CA: Physiological Function of Hypoxic Pulmonary VasoconstrictionHypoxic Pulmonary Vasoconstriction. Springer US; pp. 3–14. 2004, View Article : Google Scholar
2	K L Eales, Hollinshead KE and Tennant DA: Hypoxia and metabolic adaptation of cancer cells. Oncogenesis. 5:e1902016. View Article : Google Scholar : PubMed/NCBI
3	Vaupel P and Harrison L: Tumor hypoxia: Causative factors, compensatory mechanisms and cellular response. Oncologist. 9 Suppl 5:S4–S9. 2004. View Article : Google Scholar
4	Yamada D, Kobayashi S, Yamamoto H, Tomimaru Y, Noda T, Uemura M, Wada H, Marubashi S, Eguchi H, Tanemura M, et al: Role of the hypoxia-related gene, JMJD1A, in hepatocellular carcinoma: Clinical impact on recurrence after hepatic resection. Ann Surg Oncol. 19 Suppl 3:S3552011. View Article : Google Scholar : PubMed/NCBI
5	Andersen S, Donnem T, Al-Saad S, Al-Shibli K, Stenvold H, Busund LT and Bremnes RM: Correlation and coexpression of HIFs and NOTCH markers in NSCLC. Anticancer Res. 31:1603–1606. 2011.PubMed/NCBI
6	Coughlin SS and Ekwueme DU: Breast cancer as a global health concern. Cancer Epidemiol. 33:315–318. 2009. View Article : Google Scholar : PubMed/NCBI
7	Chakrabarti J, Turley H, Campo L, Han C, Harris AL, Gatter KC and Fox SB: The transcription factor DEC1 (stra13, SHARP2) is associated with the hypoxic response and high tumour grade in human breast cancers. Br J cancer. 91:954–958. 2004. View Article : Google Scholar : PubMed/NCBI
8	Hänze J, Eul BG, Savai R, Krick S, Goyal P, Grimminger F, Seeger W and Rose F: RNA interference for HIF-1alpha inhibits its downstream signalling and affects cellular proliferation. Biochem Biophys Res Commun. 312:571–577. 2003. View Article : Google Scholar : PubMed/NCBI
9	Zhang H, Chen J, Liu F, Gao C, Wang X, Zhao T, Liu J, Gao S, Zhao X, Ren H and Hao J: CypA, a gene downstream of HIF-1α, promotes the development of PDAC. PLoS One. 9:e928242014. View Article : Google Scholar : PubMed/NCBI
10	Teicher BA and Fricker SP: CXCL12 (SDF-1)/CXCR4 pathway in cancer. Clin Cancer Res. 16:2927–2931. 2010. View Article : Google Scholar : PubMed/NCBI
11	Huang Y, Du KM, Xue ZH, Yan H, Li D, Liu W, Chen Z, Zhao Q, Tong JH, Zhu YS and Chen GQ: Cobalt chloride and low oxygen tension trigger differentiation of acute myeloid leukemic cells: possible mediation of hypoxia-inducible factor-1alpha. Leukemia. 17:2065–2073. 2003. View Article : Google Scholar : PubMed/NCBI
12	Li S, Zhang J, Yang H, Wu C, Dang X and Liu Y: Copper depletion inhibits CoCl2-induced aggressive phenotype of MCF-7 cells via downregulation of HIF-1 and inhibition of Snail/Twist-mediated epithelial-mesenchymal transition. Sci Rep. 5:124102015. View Article : Google Scholar : PubMed/NCBI
13	Shen B, Zheng MQ, Lu JW, Jiang Q, Wang TH and Huang XE: CXCL12-CXCR4 promotes proliferation and invasion of pancreatic cancer cells. Asian Pac J Cancer Prev. 14:5403–5408. 2013. View Article : Google Scholar : PubMed/NCBI
14	Luo HQ, Xu M, Zhong WT, Cui ZY, Liu FM, Zhou KY and Li XY: EGCG decreases the expression of HIF-1α and VEGF and cell growth in MCF-7 breast cancer cells. J BUON. 19:435–439. 2014.PubMed/NCBI
15	Livak KJ and Schmittgen TD: Analysis of relative gene expression data usingreal-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
16	Semenza GL: The hypoxic tumor microenvironment: A driving force for breast cancer progression. Biochim Biophys Acta. 1863:382–391. 2016. View Article : Google Scholar : PubMed/NCBI
17	Tsutsui S, Matsuyama A, Yamamoto M, Takeuchi H, Oshiro Y, Ishida T and Maehara Y: The Akt expression correlates with the VEGF-A and -C expression as well as the microvessel and lymphatic vessel density in breast cancer. Oncol Rep. 23:621–630. 2010. View Article : Google Scholar : PubMed/NCBI
18	Zhang Z, Wu J, Liang B and Huangfu CS: Correlationbetween hypoxia inducible factor (HIF-1α) and epithelium-mesenchyma transform of ductal carcinoma with invasive breast cancer. Shandong Med J. 52:90–92. 2012.
19	Moser C, Lang SA, Mori A, Hellerbrand C, Schlitt HJ, Geissler EK, Fogler WE and Stoeltzing O: ENMD-1198, a novel tubulin binding agent rdduces HIF-lalpha and STAT3 activity in human hepatocellular carcinoma (HCC) cells and inhibits growth and vascularization in vivo. BMC Cancer. 8:2062008. View Article : Google Scholar : PubMed/NCBI
20	Marmot MG, Altman DG, Cameron DA, Dewar JA, Thompson SG and Wilcox M: The benefits and harms of breast cancer screening: An independent review. Br J Cancer. 108:2205–2240. 2013. View Article : Google Scholar : PubMed/NCBI
21	Jemal A, Bray F, Center MM, Ferlay J, Ward E and Forman D: Global cancer statistics. CA Cancer J Clin. 61:69–90. 2011. View Article : Google Scholar : PubMed/NCBI
22	Lech R and Przemyslaw O: Epidemiological models for breast cancer risk estimation. Ginekol Pol. 82:451–454. 2011.PubMed/NCBI
23	Liu ZJ, Semenza GL and Zhang HF: Hypoxia-inducible factor 1 and breast cancer metastasis. J Zhejiang Univ Sci B. 16:32–43. 2015.(In Chinese). View Article : Google Scholar : PubMed/NCBI
24	Lundgren K, Holm C and Landberg G: Hypoxia and breast cancer: Prognostic and therapeutic implications. Cell Mol Life Sci. 64:3233–3247. 2007. View Article : Google Scholar : PubMed/NCBI
25	Park SJ, Kim JG, Kim ND, Yang K, Shim JW and Heo K: Estradiol, TGF-β1 and hypoxia promote breast cancer stemness and EMT-mediated breast cancer migration. Oncol Lett. 11:1895–1902. 2016.PubMed/NCBI
26	Bradbury J: Breathing hard to keep up with HIF-1. Lancet. 358:17042001. View Article : Google Scholar : PubMed/NCBI
27	Cancer Genome Atlas Network, . Comprehensive molecular portraits of human breast tumours. Nature. 490:61–70. 2012. View Article : Google Scholar : PubMed/NCBI
28	Generali D, Berruti A, Brizzi MP, Campo L, Bonardi S, Wigfield S, Bersiga A, Allevi G, Milani M, Aguggini S, et al: Hypoxiainducible factor-1alpha expression predicts a poor response to primary chemoendocrine therapy and disease-free survival in primary human breast cancer. Clin Cancer Res. 12:4562–4568. 2006. View Article : Google Scholar : PubMed/NCBI
29	Gruber G, Greiner RH, Hlushchuk R, Aebersold DM, Altermatt HJ, Berclaz G and Djonov V: Hypoxia-inducible factor 1alpha in high-risk breast cancer: An independent prognostic parameter. Breast Cancer Res. 6:R191–R198. 2004. View Article : Google Scholar : PubMed/NCBI
30	Goswami S, Sahai E, Wyckoff JB, Cammer M, Cox D, Pixley FJ, Stanley ER, Segall JE and Condeelis JS: Macrophages promote the invasion of breast carcinoma cells via a colony-stimulating factor-1/epidermal growth factor paracrine loop. Cancer Res. 65:5278–5283. 2005. View Article : Google Scholar : PubMed/NCBI
31	Rahimi M, Toth TA and Tang CK: CXCR4 suppression attenuates EGFRVIII mediated invasion and induces p38 MAPK-dependent protein trafficking and degradation of EGFRvIII in breast cancer cells. Cancer Lett. 306:43–51. 2011. View Article : Google Scholar : PubMed/NCBI
32	Helbig G, Christopherson KW II, Bhat-Nakshatfi P, Kumar S, Kishimoto H, Miller KD, Broxmeyer HE and Nakshatri H: NF-kappaB promotes breast cancer cell migration and metastasis by inducing the expression of the chemokine receptor CXCR4. J Biol Chem. 278:21631–21638. 2003. View Article : Google Scholar : PubMed/NCBI
33	Parker CC, Kim RH, Li BD and Chu QD: The chemokine receptor CXCR4 as a novel independent prognosic marker for node-positive breast cancer patients. J Surg Oncol. 106:393–398. 2012. View Article : Google Scholar : PubMed/NCBI
34	Dunn LK, Mohammad KS, Fournier PG, McKenna CR, Davis HW, Niewolna M, Peng XH, Chirgwin JM and Guise TA: Hypoxia and TGF-beta drive breast cancer bone metastases through parallel signaling pathways in tumor cells and the bone microenvironment. PLoS One. 4:e68962009. View Article : Google Scholar : PubMed/NCBI
35	Citti A, Boldrini R, Inserra A, Alisi A, Pessolano R, Mastronuzzi A, Zin A, De Sio L, Rosolen A, Locatelli F and Fruci D: Expression of multidrug resistance-associated proteins in paediatric soft tissue sarcomas before and after chemotherapy. Int J Oncol. 41:117–124. 2012.PubMed/NCBI
36	Mego M, Cholujova D, Minarik G, Sedlackova T, Gronesova P, Karaba M, Benca J, Cingelova S, Cierna Z, Manasova D, et al: CXCR4-SDF-1 interaction potentially mediates trafficking of circulating tumor cells in primary breast cancer. Bmc Cancer. 16:1272016. View Article : Google Scholar : PubMed/NCBI
37	Ferrara N: VEGF and the quest for tumour angiogenesis factors. Nat Rev Cancer. 2:795–803. 2002. View Article : Google Scholar : PubMed/NCBI
38	Timoshenko AV, Chakraborty C, Wagner GF and Lala PK: COX-2-mediated stimulation of the lymphangiogenic factor VEGF-C in human breast cancer. Br J Cancer. 94:1154–1163. 2006. View Article : Google Scholar : PubMed/NCBI
39	Komatsu DE and Hadjiargyrou M: Activation of the transcription factor HIF-1 and its target genes, VEGF, HO-1, iNOS, during fracture repair. Bone. 34:680–688. 2004. View Article : Google Scholar : PubMed/NCBI
40	Semenza GL: Expression of hypoxia-inducible factor 1: Mechanisms and consequences. Biochem Pharmacol. 59:47–53. 2000. View Article : Google Scholar : PubMed/NCBI
41	Li G, He L, Zhang E, Shi J, Zhang Q, Le AD, Zhou K and Tang X: Overexpression of human papillomavirus (HPV) type 16 oncoproteins promotes angiogenesis via enhancing HIF-1α and VEGF expression in non-small cell lung cancer cells. Cancer Lett. 311:160–170. 2011. View Article : Google Scholar : PubMed/NCBI
42	Dong X, Wang YS, Dou GR, Hou HY, Shi YY, Zhang R, Ma K, Wu L, Yao LB, Cai Y and Zhang J: Influence of Dll4 via HIF-1α-VEGF signaling on the angiogenesis of choroidal neovascularization under hypoxic conditions. PLoS One. 6:e184812011. View Article : Google Scholar : PubMed/NCBI
43	Ward C, Langdon SP, Mullen P, Harris AL, Harrison DJ, Supuran CT and Kunkler IH: New strategies for targeting the hypoxic tumour microenvironment in breast cancer. Cancer Treat Rev. 39:171–179. 2013.(In Chinese). View Article : Google Scholar : PubMed/NCBI
44	Lopez-Haber C, Barrio-Real L, Casado-Medrano V and Kazanietz MG: Heregulin/ErbB3 signaling enhances CXCR4-driven rac1 activation and breast cancer cell motility via hypoxia-inducible factor 1α. Mol Cell Biol. 36:2011–2026. 2016. View Article : Google Scholar : PubMed/NCBI