2-Methoxyestradiol enhances radiosensitivity in radioresistant melanoma MDA-MB-435R cells by regulating glycolysis via HIF-1α/PDK1 axis

Zhao,Hong; Jiang,Huangang; Li,Zheng; Zhuang,Yafei; Liu,Yinyin; Zhou,Shuliang; Xiao,Youde; Xie,Conghua; Zhou,Fuxiang; Zhou,Yunfeng

doi:10.3892/ijo.2017.3924

2-Methoxyestradiol enhances radiosensitivity in radioresistant melanoma MDA-MB-435R cells by regulating glycolysis via HIF-1α/PDK1 axis

Authors:
- Hong Zhao
- Huangang Jiang
- Zheng Li
- Yafei Zhuang
- Yinyin Liu
- Shuliang Zhou
- Youde Xiao
- Conghua Xie
- Fuxiang Zhou
- Yunfeng Zhou
View Affiliations

Affiliations: Hubei Key Laboratory of Tumor Biological Behaviors, Zhongnan Hospital of Wuhan University, Wuchang, Wuhan, Hubei 430071, P.R. China, Department of Radiation and Medical Oncology, Zhongnan Hospital of Wuhan University, Wuchang, Wuhan, Hubei 430071, P.R. China
Published online on: March 22, 2017 https://doi.org/10.3892/ijo.2017.3924
Pages: 1531-1540
Copyright: © Zhao et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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

HIF-1α overexpression is associated with radioresistance of various cancers. A radioresistant human melanoma cell model MDA-MB-435R (435R) was established by us previously. Compared with the parental cells MDA-MB‑435 (435S), an elevated level of HIF-1α expression in 435R cells was demonstrated in our recent experiments. Therefore, in the current study, we sought to determine whether selective HIF-1α inhibitors could radiosensitize the 435R cells to X-ray, and to identify the potential mechanisms. Our data demonstrated that inhibition of HIF-1α with 2-methoxyestradiol (2-MeOE2) significantly enhanced radiosensitivity of 435R cells. 2-MeOE2 increased DNA damage and ratio of apoptosis cells induced by irradiation. Whereas, cell proliferation and the expression of pyruvate dehydrogenase kinase 1 (PDK1) were decreased after 2-MeOE2 treatment. The change of expression of GLUT1, LDHA and the cellular ATP level and extracellular lactate production indicates that 2-MeOE2 suppressed glycolytic state of 435R cells. In addition, the radioresistance, glycolytic state and cell proliferation of 435R cells were also decreased after inhibiting pyruvate dehydrogenase kinase 1 (PDK1) with dichloroacetate (DCA). DCA could also increase DNA damage and ratio of apoptotic cells induced by irradiation. These results also suggest that inhibition of HIF-1α with 2-MeOE2 sensitizes radioresistant melanoma cells 435R to X-ray irradiation through targeting the glycolysis that is regulated by PDK1. Selective inhibitors of HIF-1α and glycolysis are potential drugs to enhance radiosensitivity of melanoma cells.

View Figures

View References

1	Slominski A, Kim TK, Brożyna AA, Janjetovic Z, Brooks DL, Schwab LP, Skobowiat C, Jóźwicki W and Seagroves TN: The role of melanogenesis in regulation of melanoma behavior: Melanogenesis leads to stimulation of HIF-1α expression and HIF-dependent attendant pathways. Arch Biochem Biophys. 563:79–93. 2014. View Article : Google Scholar : PubMed/NCBI
2	Aebersold DM, Burri P, Beer KT, Laissue J, Djonov V, Greiner RH and Semenza GL: Expression of hypoxia-inducible factor-1alpha: A novel predictive and prognostic parameter in the radiotherapy of oropharyngeal cancer. Cancer Res. 61:2911–2916. 2001.PubMed/NCBI
3	Koukourakis MI, Giatromanolaki A, Sivridis E, Simopoulos C, Turley H, Talks K, Gatter KC and Harris AL: Hypoxia-inducible factor (HIF1A and HIF2A), angiogenesis, and chemoradiotherapy outcome of squamous cell head-and-neck cancer. Int J Radiat Oncol Biol Phys. 53:1192–1202. 2002. View Article : Google Scholar : PubMed/NCBI
4	Mabjeesh NJ, Escuin D, LaVallee TM, Pribluda VS, Swartz GM, Johnson MS, Willard MT, Zhong H, Simons JW and Giannakakou P: 2ME2 inhibits tumor growth and angiogenesis by disrupting microtubules and dysregulating HIF. Cancer Cell. 3:363–375. 2003. View Article : Google Scholar : PubMed/NCBI
5	Maxwell PH, Pugh CW and Ratcliffe PJ: The pVHL-hIF-1 system. A key mediator of oxygen homeostasis. Adv Exp Med Biol. 502:365–376. 2001. View Article : Google Scholar
6	Zeng L, Zhou HY, Tang NN, Zhang WF, He GJ, Hao B, Feng YD and Zhu H: Wortmannin influences hypoxia-inducible factor-1 alpha expression and glycolysis in esophageal carcinoma cells. World J Gastroenterol. 22:4868–4880. 2016. View Article : Google Scholar : PubMed/NCBI
7	Chen Y, Cao KE, Wang S, Chen J, He B, He GU, Chen Y, Peng B and Zhou J: MicroRNA-138 suppresses proliferation, invasion and glycolysis in malignant melanoma cells by targeting HIF-1α. Exp Ther Med. 11:2513–2518. 2016.PubMed/NCBI
8	Kim JW, Tchernyshyov I, Semenza GL and Dang CV: HIF-1-mediated expression of pyruvate dehydrogenase kinase: A metabolic switch required for cellular adaptation to hypoxia. Cell Metab. 3:177–185. 2006. View Article : Google Scholar : PubMed/NCBI
9	Fujiwara S, Kawano Y, Yuki H, Okuno Y, Nosaka K, Mitsuya H and Hata H: PDK1 inhibition is a novel therapeutic target in multiple myeloma. Br J Cancer. 108:170–178. 2013.PubMed/NCBI
10	Vander Heiden MG, Cantley LC and Thompson CB: Understanding the Warburg effect: The metabolic requirements of cell proliferation. Science. 324:1029–1033. 2009. View Article : Google Scholar : PubMed/NCBI
11	Koppenol WH, Bounds PL and Dang CV: Otto Warburg's contributions to current concepts of cancer metabolism. Nat Rev Cancer. 11:325–337. 2011. View Article : Google Scholar : PubMed/NCBI
12	Warburg O: On respiratory impairment in cancer cells. Science. 124:269–270. 1956.PubMed/NCBI
13	Scott DA, Richardson AD, Filipp FV, Knutzen CA, Chiang GG, Ronai ZA, Osterman AL and Smith JW: Comparative metabolic flux profiling of melanoma cell lines: Beyond the Warburg effect. J Biol Chem. 286:42626–42634. 2011. View Article : Google Scholar : PubMed/NCBI
14	Bettum IJ, Gorad SS, Barkovskaya A, Pettersen S, Moestue SA, Vasiliauskaite K, Tenstad E, Øyjord T, Risa Ø, Nygaard V, et al: Metabolic reprogramming supports the invasive phenotype in malignant melanoma. Cancer Lett. 366:71–83. 2015. View Article : Google Scholar : PubMed/NCBI
15	Bhatt AN, Chauhan A, Khanna S, Rai Y, Singh S, Soni R, Kalra N and Dwarakanath BS: Transient elevation of glycolysis confers radio-resistance by facilitating DNA repair in cells. BMC Cancer. 15:3352015. View Article : Google Scholar : PubMed/NCBI
16	Zhong J, Rajaram N, Brizel DM, Frees AE, Ramanujam N, Batinic-Haberle I and Dewhirst MW: Radiation induces aerobic glycolysis through reactive oxygen species. Radiother Oncol. 106:390–396. 2013. View Article : Google Scholar : PubMed/NCBI
17	Shimura T, Noma N, Sano Y, Ochiai Y, Oikawa T, Fukumoto M and Kunugita N: AKT-mediated enhanced aerobic glycolysis causes acquired radioresistance by human tumor cells. Radiother Oncol. 112:302–307. 2014. View Article : Google Scholar : PubMed/NCBI
18	Yang C, Qin Y, Zhang H, Ma J and Yang K: Upregulation of SLC19A2 in stem cell-like cancer cells of nasopharyngeal carcinoma contributes to increased radioresistance through enhanced glycolysis. Int J Radiat Oncol Biol Phys. 96:E5822016. View Article : Google Scholar
19	Jiang S, Wang R, Yan H, Jin L, Dou X and Chen D: MicroRNA-21 modulates radiation resistance through upregulation of hypoxia-inducible factor-1α-promoted glycolysis in non-small cell lung cancer cells. Mol Med Rep. 13:4101–4107. 2016.PubMed/NCBI
20	Liu G, Li YI and Gao X: Overexpression of microRNA-133b sensitizes non-small cell lung cancer cells to irradiation through the inhibition of glycolysis. Oncol Lett. 11:2903–2908. 2016.PubMed/NCBI
21	Shen H, Hau E, Joshi S, Dilda PJ and McDonald KL: Sensitization of glioblastoma cells to irradiation by modulating the glucose metabolism. Mol Cancer Ther. 14:1794–1804. 2015. View Article : Google Scholar : PubMed/NCBI
22	Luo YM, Xia NX, Yang L, Li Z, Yang H, Yu HJ, Liu Y, Lei H, Zhou FX, Xie CH, et al: CTC1 increases the radioresistance of human melanoma cells by inhibiting telomere shortening and apoptosis. Int J Mol Med. 33:1484–1490. 2014.PubMed/NCBI
23	Franken NA, Rodermond HM, Stap J, Haveman J and van Bree C: Clonogenic assay of cells in vitro. Nat Protoc. 1:2315–2319. 2006. View Article : Google Scholar
24	Rafehi H, Orlowski C, Georgiadis GT, Ververis K, El-Osta A and Karagiannis TC: Clonogenic assay: Adherent cells. J Vis Exp. 49:25732011.
25	Ning S, Shui C, Khan WB, Benson W, Lacey DL and Knox SJ: Effects of keratinocyte growth factor on the proliferation and radiation survival of human squamous cell carcinoma cell lines in vitro and in vivo. Int J Radiat Oncol Biol Phys. 40:177–187. 1998. View Article : Google Scholar : PubMed/NCBI
26	Rao PN, Cessac JW, Tinley TL and Mooberry SL: Synthesis and antimitotic activity of novel 2-methoxyestradiol analogs. Steroids. 67:1079–1089. 2002. View Article : Google Scholar : PubMed/NCBI
27	Hu D and Kipps TJ: Reduction in mitochondrial membrane potential is an early event in Fas-independent CTL-mediated apoptosis. Cell Immunol. 195:43–52. 1999. View Article : Google Scholar : PubMed/NCBI
28	Jenrette JM: Malignant melanoma: The role of radiation therapy revisited. Semin Oncol. 23:759–762. 1996.PubMed/NCBI
29	Stevens G and McKay MJ: Dispelling the myths surrounding radiotherapy for treatment of cutaneous melanoma. Lancet Oncol. 7:575–583. 2006. View Article : Google Scholar : PubMed/NCBI
30	Rofstad EK: Radiation biology of malignant melanoma. Acta Radiol Oncol. 25:1–10. 1986. View Article : Google Scholar : PubMed/NCBI
31	Overgaard J, Overgaard M, Hansen PV and von der Maase H: Some factors of importance in the radiation treatment of malignant melanoma. Radiother Oncol. 5:183–192. 1986. View Article : Google Scholar : PubMed/NCBI
32	Masoud GN and Li W: HIF-1α pathway: Role, regulation and intervention for cancer therapy. Acta Pharm Sin B. 5:378–389. 2015. View Article : Google Scholar : PubMed/NCBI
33	Pribluda VS, Gubish ER Jr, Lavallee TM, Treston A, Swartz GM and Green SJ: 2-Methoxyestradiol: An endogenous antiangiogenic and antiproliferative drug candidate. Cancer Metastasis Rev. 19:173–179. 2000. View Article : Google Scholar
34	James J, Murry DJ, Treston AM, Storniolo AM, Sledge GW, Sidor C and Miller KD: Phase I safety, pharmacokinetic and pharmacodynamic studies of 2-methoxyestradiol alone or in combination with docetaxel in patients with locally recurrent or metastatic breast cancer. Invest New Drugs. 25:41–48. 2007. View Article : Google Scholar
35	Bruce JY, Eickhoff J, Pili R, Logan T, Carducci M, Arnott J, Treston A, Wilding G and Liu G: A phase II study of 2-methoxyestradiol nanocrystal colloidal dispersion alone and in combination with sunitinib malate in patients with metastatic renal cell carcinoma progressing on sunitinib malate. Invest New Drugs. 30:794–802. 2012. View Article : Google Scholar
36	Sweeney C, Liu G, Yiannoutsos C, Kolesar J, Horvath D, Staab MJ, Fife K, Armstrong V, Treston A, Sidor C, et al: A phase II multicenter, randomized, double-blind, safety trial assessing the pharmacokinetics, pharmacodynamics, and efficacy of oral 2-methoxyestradiol capsules in hormone-refractory prostate cancer. Clin Cancer Res. 11:6625–6633. 2005. View Article : Google Scholar : PubMed/NCBI
37	Xiao X, Huang X, Ye F, Chen B, Song C, Wen J, Zhang Z, Zheng G, Tang H and Xie X: The miR-34a-LDHA axis regulates glucose metabolism and tumor growth in breast cancer. Sci Rep. 6:217352016. View Article : Google Scholar : PubMed/NCBI
38	Zhao D, Zou SW, Liu Y, Zhou X, Mo Y, Wang P, Xu YH, Dong B, Xiong Y, Lei QY, et al: Lysine-5 acetylation negatively regulates lactate dehydrogenase A and is decreased in pancreatic cancer. Cancer Cell. 23:464–476. 2013. View Article : Google Scholar : PubMed/NCBI
39	Shi M, Cui J, Du J, Wei D, Jia Z, Zhang J, Zhu Z, Gao Y and Xie K: A novel KLF4/LDHA signaling pathway regulates aerobic glycolysis in and progression of pancreatic cancer. Clin Cancer Res. 20:4370–4380. 2014. View Article : Google Scholar : PubMed/NCBI
40	Shibuya K, Okada M, Suzuki S, Seino M, Seino S, Takeda H and Kitanaka C: Targeting the facilitative glucose transporter GLUT1 inhibits the self-renewal and tumor-initiating capacity of cancer stem cells. Oncotarget. 6:651–661. 2015. View Article : Google Scholar :
41	Hoeijmakers JH: Genome maintenance mechanisms for preventing cancer. Nature. 411:366–374. 2001. View Article : Google Scholar : PubMed/NCBI
42	Lips J and Kaina B: DNA double-strand breaks trigger apoptosis in p53-deficient fibroblasts. Carcinogenesis. 22:579–585. 2001. View Article : Google Scholar : PubMed/NCBI
43	Fortini P, Ferretti C and Dogliotti E: The response to DNA damage during differentiation: Pathways and consequences. Mutat Res. 743–744:160–168. 2013. View Article : Google Scholar
44	Zou H, Zhao S, Zhang J, Lv G, Zhang X, Yu H, Wang H and Wang L: Enhanced radiation-induced cytotoxic effect by 2-ME in glioma cells is mediated by induction of cell cycle arrest and DNA damage via activation of ATM pathways. Brain Res. 1185:231–238. 2007. View Article : Google Scholar : PubMed/NCBI
45	Pierce AJ, Hu P, Han M, Ellis N and Jasin M: Ku DNA end-binding protein modulates homologous repair of double-strand breaks in mammalian cells. Genes Dev. 15:3237–3242. 2001. View Article : Google Scholar : PubMed/NCBI
46	Little JB, Hahn GM, Frindel E and Tubiana M: Repair of potentially lethal radiation damage in vitro and in vivo. Radiology. 106:689–694. 1973. View Article : Google Scholar : PubMed/NCBI
47	Mueck AO and Seeger H: 2-Methoxyestradiol - biology and mechanism of action. Steroids. 75:625–631. 2010. View Article : Google Scholar : PubMed/NCBI
48	Long F, Si L, Long X, Yang B, Wang X and Zhang F: 2ME2 increase radiation-induced apoptosis of keloid fibroblasts by targeting HIF-1α in vitro. Australas J Dermatol. 57:e32–e38. 2016. View Article : Google Scholar
49	Aquino-Gálvez A, González-Ávila G, Delgado-Tello J, Castillejos-López M, Mendoza-Milla C, Zúñiga J, Checa M, Maldonado-Martínez HA, Trinidad-López A, Cisneros J, et al: Effects of 2-methoxyestradiol on apoptosis and HIF-1α and HIF-2α expression in lung cancer cells under normoxia and hypoxia. Oncol Rep. 35:577–583. 2016.