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Open Access 01-12-2014 | Research

MicroRNA-566 activates EGFR signaling and its inhibition sensitizes glioblastoma cells to nimotuzumab

Authors: Kai-Liang Zhang, Xuan Zhou, Lei Han, Lu-Yue Chen, Ling-Chao Chen, Zhen-Dong Shi, Ming Yang, Yu Ren, Jing-Xuan Yang, Thomas S Frank, Chuan-Bao Zhang, Jun-Xia Zhang, Pei-Yu Pu, Jian-Ning Zhang, Tao Jiang, Eric J Wagner, Min Li, Chun-Sheng Kang

Published in: Molecular Cancer | Issue 1/2014

Abstract

Background

Epidermal growth factor receptor (EGFR) is amplified in 40% of human glioblastomas. However, most glioblastoma patients respond poorly to anti-EGFR therapy. MicroRNAs can function as either oncogenes or tumor suppressor genes, and have been shown to play an important role in cancer cell proliferation, invasion and apoptosis. Whether microRNAs can impact the therapeutic effects of EGFR inhibitors in glioblastoma is unknown.

Methods

miR-566 expression levels were detected in glioma cell lines, using real-time quantitative RT-PCR (qRT-PCR). Luciferase reporter assays and Western blots were used to validate VHL as a direct target gene of miR-566. Cell proliferation, invasion, cell cycle distribution and apoptosis were also examined to confirm whether miR-566 inhibition could sensitize anti-EGFR therapy.

Results

In this study, we demonstrated that miR-566 is up-regulated in human glioma cell lines and inhibition of miR-566 decreased the activity of the EGFR pathway. Lentiviral mediated inhibition of miR-566 in glioblastoma cell lines significantly inhibited cell proliferation and invasion and led to cell cycle arrest in the G₀/G₁ phase. In addition, we identified von Hippel-Lindau (VHL) as a novel functional target of miR-566. VHL regulates the formation of the β-catenin/hypoxia-inducible factors-1α complex under miR-566 regulation.

Conclusions

miR-566 activated EGFR signaling and its inhibition sensitized glioblastoma cells to anti-EGFR therapy.

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Lacroix M, Abi-Said D, Fourney DR, Gokaslan ZL, Shi W, DeMonte F, Lang FF, McCutcheon IE, Hassenbusch SJ, Holland E, Hess K, Michael C, Miller D, Sawaya R: A multivariate analysis of 416 patients with glioblastoma multiforme: prognosis, extent of resection, and survival. Journal of Neurosurgery. 2001, 95: 190-198. 10.3171/jns.2001.95.2.0190.CrossRefPubMed

Rich JN, Hans C, Jones B, Iversen ES, McLendon RE, Rasheed BK, Dobra A, Dressman HK, Bigner DD, Nevins JR, West M: Gene expression profiling and genetic markers in glioblastoma survival. Cancer Res. 2005, 65: 4051-4058. 10.1158/0008-5472.CAN-04-3936CrossRefPubMed

Zhu VF, Yang J, Lebrun DG, Li M: Understanding the role of cytokines in Glioblastoma Multiforme pathogenesis. Cancer Lett. 2012, 316: 139-150. 10.1016/j.canlet.2011.11.001CrossRefPubMed

Smith JS, Tachibana I, Passe SM, Huntley BK, Borell TJ, Iturria N, O'Fallon JR, Schaefer PL, Scheithauer BW, James CD, Buckner JC, Jenkins RB: PTEN mutation, EGFR amplification, and outcome in patients with anaplastic astrocytoma and glioblastoma multiforme. J Natl Cancer Inst. 2001, 93: 1246-1256. 10.1093/jnci/93.16.1246CrossRefPubMed

Lynch TJ, Bell DW, Sordella R, Gurubhagavatula S, Okimoto RA, Brannigan BW, Harris PL, Haserlat SM, Supko JG, Haluska FG, Louis DN, Christiani DC, Settleman J, Haber DA: Activating mutations in the epidermal growth factor receptor underlying responsiveness of non-small-cell lung cancer to gefitinib. N Engl J Med. 2004, 350: 2129-2139. 10.1056/NEJMoa040938CrossRefPubMed

Chakravarti A, Chakladar A, Delaney MA, Latham DE, Loeffler JS: The epidermal growth factor receptor pathway mediates resistance to sequential administration of radiation and chemotherapy in primary human glioblastoma cells in a RAS-dependent manner. Cancer Res. 2002, 62: 4307-4315.PubMed

Schulte JH, Horn S, Schlierf S, Schramm A, Heukamp LC, Christiansen H, Buettner R, Berwanger B, Eggert A: MicroRNAs in the pathogenesis of neuroblastoma. Cancer Lett. 2009, 274: 10-15. 10.1016/j.canlet.2008.06.010CrossRefPubMed

Zhou X, Ren Y, Moore L, Mei M, You Y, Xu P, Wang B, Wang G, Jia Z, Pu P, Zhang W, Kang C: Downregulation of miR-21 inhibits EGFR pathway and suppresses the growth of human glioblastoma cells independent of PTEN status. Lab Invest. 2010, 90: 144-155. 10.1038/labinvest.2009.126CrossRefPubMed

Chen L, Han L, Zhang K, Shi Z, Zhang J, Zhang A, Wang Y, Song Y, Li Y, Jiang T, Pu P, Jiang C, Kang C: VHL regulates the effects of miR-23b on glioma survival and invasion via suppression of HIF-1alpha/VEGF and beta-catenin/Tcf-4 signaling. Neuro Oncol. 2012, 14: 1026-1036. 10.1093/neuonc/nos122PubMedCentralCrossRefPubMed

10.

Chen L, Li H, Han L, Zhang K, Wang G, Wang Y, Liu Y, Zheng Y, Jiang T, Pu P, Jiang C, Kang C: Expression and function of miR-27b in human glioma. Oncol Rep. 2011, 26: 1617-1621.PubMed

11.

Chen L, Zhang W, Yan W, Han L, Zhang K, Shi Z, Zhang J, Wang Y, Li Y, Yu S, Pu P, Jiang C, Jiang T, Kang C: The putative tumor suppressor miR-524-5p directly targets Jagged-1 and Hes-1 in glioma. Carcinogenesis. 2012, 33: 2276-2282. 10.1093/carcin/bgs261CrossRefPubMed

12.

Zhang W, Zhang J, Yan W, You G, Bao Z, Li S, Kang C, Jiang C, You Y, Zhang Y, Chen CC, Song SW, Jiang T: Whole-genome microRNA expression profiling identifies a 5-microRNA signature as a prognostic biomarker in Chinese patients with primary glioblastoma multiforme. Cancer. 2013, 119: 814-824. 10.1002/cncr.27826CrossRefPubMed

13.

Wang XF, Shi ZM, Wang XR, Cao L, Wang YY, Zhang JX, Yin Y, Luo H, Kang CS, Liu N, Jiang T, You YP: MiR-181d acts as a tumor suppressor in glioma by targeting K-ras and Bcl-2. J Cancer Res Clin Oncol. 2012, 138: 573-584. 10.1007/s00432-011-1114-xCrossRefPubMed

14.

Baffa R, Fassan M, Volinia S, O’Hara B, Liu CG, Palazzo JP, Gardiman M, Rugge M, Gomella LG, Croce CM, Rosenberg A: MicroRNA expression profiling of human metastatic cancers identifies cancer gene targets. J Pathol. 2009, 219: 214-221. 10.1002/path.2586CrossRefPubMed

15.

Dudziec E, Miah S, Choudhry HM, Owen HC, Blizard S, Glover M, Hamdy FC, Catto JW: Hypermethylation of CpG islands and shores around specific microRNAs and mirtrons is associated with the phenotype and presence of bladder cancer. Clin Cancer Res. 2011, 17: 1287-1296. 10.1158/1078-0432.CCR-10-2017CrossRefPubMed

16.

Sarkar FH, Li Y, Wang Z, Kong D, Ali S: Implication of microRNAs in drug resistance for designing novel cancer therapy. Drug Resist Updat. 2010, 13: 57-66. 10.1016/j.drup.2010.02.001PubMedCentralCrossRefPubMed

17.

Weidhaas JB, Babar I, Nallur SM, Trang P, Roush S, Boehm M, Gillespie E, Slack FJ: MicroRNAs as potential agents to alter resistance to cytotoxic anticancer therapy. Cancer Res. 2007, 67: 11111-11116. 10.1158/0008-5472.CAN-07-2858CrossRefPubMed

18.

Zhang JX, Qian D, Wang FW, Liao DZ, Wei JH, Tong ZT, Fu J, Huang XX, Liao YJ, Deng HX, Zeng YX, Xie D, Mai SJ: MicroRNA-29c enhances the sensitivities of human nasopharyngeal carcinoma to cisplatin-based chemotherapy and radiotherapy. Cancer Lett. 2013, 329: 91-98. 10.1016/j.canlet.2012.10.033CrossRefPubMed

19.

Weiss GJ, Bemis LT, Nakajima E, Sugita M, Birks DK, Robinson WA, Varella-Garcia M, Bunn PA, Haney J, Helfrich BA, Kato H, Hirsch FR, Franklin WA: EGFR regulation by microRNA in lung cancer: correlation with clinical response and survival to gefitinib and EGFR expression in cell lines. Ann Oncol. 2008, 19: 1053-1059. 10.1093/annonc/mdn006CrossRefPubMed

20.

Papagiannakopoulos T, Shapiro A, Kosik KS: MicroRNA-21 targets a network of key tumor-suppressive pathways in glioblastoma cells. Cancer Res. 2008, 68: 8164-8172. 10.1158/0008-5472.CAN-08-1305CrossRefPubMed

21.

Kanno H, Sato H, Yokoyama TA, Yoshizumi T, Yamada S: The VHL tumor suppressor protein regulates tumorigenicity of U87-derived glioma stem-like cells by inhibiting the JAK/STAT signaling pathway. Int J Oncol. 2013, 42: 881-886.PubMed

22.

Behrens J: One hit, two outcomes for VHL-mediated tumorigenesis. Nat Cell Biol. 2008, 10: 1127-1128. 10.1038/ncb1008-1127CrossRefPubMed

23.

Zhang K, Zhang J, Han L, Pu P, Kang C: Wnt/beta-catenin signaling in glioma. J Neuroimmune Pharmacol. 2012, 7: 740-749. 10.1007/s11481-012-9359-yCrossRefPubMed

24.

Grimaldi AM, Guida T, D’Attino R, Perrotta E, Otero M, Masala A, Carteni G: Sunitinib: bridging present and future cancer treatment. Ann Oncol. 2007, 18 (Suppl 6): vi31-34.PubMed

25.

Maemondo M, Inoue A, Kobayashi K, Sugawara S, Oizumi S, Isobe H, Gemma A, Harada M, Yoshizawa H, Kinoshita I, Fujita Y, Okinaga S, Hirano H, Yoshimori K, Harada T, Ogura T, Ando M, Miyazawa H, Tanaka T, Saijo Y, Hagiwara K, Morita S, Nukiwa T: Gefitinib or chemotherapy for non-small-cell lung cancer with mutated EGFR. N Engl J Med. 2010, 362: 2380-2388. 10.1056/NEJMoa0909530CrossRefPubMed

26.

Zhu H, Cao X, Ali-Osman F, Keir S, Lo HW: EGFR and EGFRvIII interact with PUMA to inhibit mitochondrial translocalization of PUMA and PUMA-mediated apoptosis independent of EGFR kinase activity. Cancer Lett. 2010, 294: 101-110.PubMedCentralCrossRefPubMed

27.

Nautiyal J, Majumder P, Patel BB, Lee FY, Majumdar AP: Src inhibitor dasatinib inhibits growth of breast cancer cells by modulating EGFR signaling. Cancer Lett. 2009, 283: 143-151. 10.1016/j.canlet.2009.03.035CrossRefPubMed

28.

Dienstmann R, De Dosso S, Felip E, Tabernero J: Drug development to overcome resistance to EGFR inhibitors in lung and colorectal cancer. Mol Oncol. 2012, 6: 15-26. 10.1016/j.molonc.2011.11.009CrossRefPubMed

29.

Lo HW: EGFR-targeted therapy in malignant glioma: novel aspects and mechanisms of drug resistance. Curr Mol Pharmacol. 2010, 3: 37-52. 10.2174/1874467211003010037PubMedCentralCrossRefPubMed

30.

Murad JP, Lin OA, Espinosa EV, Khasawneh FT: Current and experimental antibody-based therapeutics: insights, breakthroughs, setbacks and future directions. Curr Mol Med. 2013, 13: 165-178. 10.2174/156652413804486322CrossRefPubMed

31.

Engelman JA, Janne PA: Mechanisms of acquired resistance to epidermal growth factor receptor tyrosine kinase inhibitors in non-small cell lung cancer. Clin Cancer Res. 2008, 14: 2895-2899. 10.1158/1078-0432.CCR-07-2248CrossRefPubMed

32.

Hara F, Aoe M, Doihara H, Taira N, Shien T, Takahashi H, Yoshitomi S, Tsukuda K, Toyooka S, Ohta T, Shimizu N: Antitumor effect of gefitinib (‘Iressa’) on esophageal squamous cell carcinoma cell lines in vitro and in vivo. Cancer Lett. 2005, 226: 37-47. 10.1016/j.canlet.2004.12.025CrossRefPubMed

33.

Mellinghoff IK, Wang MY, Vivanco I, Haas-Kogan DA, Zhu S, Dia EQ, Lu KV, Yoshimoto K, Huang JH, Chute DJ, Riggs BL, Horvath S, Liau LM, Cavenee WK, Rao PN, Beroukhim R, Peck TC, Lee JC, Sellers WR, Stokoe D, Prados M, Cloughesy TF, Sawyers CL, Mischel PS: Molecular determinants of the response of glioblastomas to EGFR kinase inhibitors. N Engl J Med. 2005, 353: 2012-2024. 10.1056/NEJMoa051918CrossRefPubMed

34.

Osio A, Mateus C, Soria JC, Massard C, Malka D, Boige V, Besse B, Robert C: Cutaneous side-effects in patients on long-term treatment with epidermal growth factor receptor inhibitors. Br J Dermatol. 2009, 161: 515-521. 10.1111/j.1365-2133.2009.09214.xCrossRefPubMed

35.

Crombet T, Osorio M, Cruz T, Roca C, del Castillo R, Mon R, Iznaga-Escobar N, Figueredo R, Koropatnick J, Renginfo E, Fernandez E, Alvarez D, Torres O, Ramos M, Leonard I, Perez R, Lage A: Use of the humanized anti-epidermal growth factor receptor monoclonal antibody h-R3 in combination with radiotherapy in the treatment of locally advanced head and neck cancer patients. J Clin Oncol. 2004, 22: 1646-1654. 10.1200/JCO.2004.03.089CrossRefPubMed

36.

Kefas B, Godlewski J, Comeau L, Li Y, Abounader R, Hawkinson M, Lee J, Fine H, Chiocca EA, Lawler S: Purow B: microRNA-7 inhibits the epidermal growth factor receptor and the Akt pathway and is down-regulated in glioblastoma. Cancer Res. 2008, 68: 3566-3572. 10.1158/0008-5472.CAN-07-6639CrossRefPubMed

37.

Sorrentino A, Liu CG, Addario A, Peschle C, Scambia G, Ferlini C: Role of microRNAs in drug-resistant ovarian cancer cells. Gynecol Oncol. 2008, 111: 478-486. 10.1016/j.ygyno.2008.08.017CrossRefPubMed

38.

Graziano F, Canestrari E, Loupakis F, Ruzzo A, Galluccio N, Santini D, Rocchi M, Vincenzi B, Salvatore L, Cremolini C, Spoto C, Catalano V, D'Emidio S, Giordani P, Tonini G, Falcone A, Magnani M: Genetic modulation of the Let-7 microRNA binding to KRAS 3'-untranslated region and survival of metastatic colorectal cancer patients treated with salvage cetuximab-irinotecan. Pharmacogenomics J. 2010, 10: 458-464. 10.1038/tpj.2010.9CrossRefPubMed

39.

Slaby O, Lakomy R, Fadrus P, Hrstka R, Kren L, Lzicarova E, Smrcka M, Svoboda M, Dolezalova H, Novakova J, Valik D, Vyzula R, Michalek J: MicroRNA-181 family predicts response to concomitant chemoradiotherapy with temozolomide in glioblastoma patients. Neoplasma. 2010, 57: 264-269. 10.4149/neo_2010_03_264CrossRefPubMed

40.

Auffinger B, Thaci B, Ahmed A, Ulasov I, Lesniak MS: MicroRNA targeting as a therapeutic strategy against glioma. Curr Mol Med. 2013, 13: 535-542. 10.2174/1566524011313040006CrossRefPubMed

41.

Adam L, Zhong M, Choi W, Qi W, Nicoloso M, Arora A, Calin G, Wang H, Siefker-Radtke A, McConkey D, Bar-Eli M, Dinney C: miR-200 expression regulates epithelial-to-mesenchymal transition in bladder cancer cells and reverses resistance to epidermal growth factor receptor therapy. Clin Cancer Res. 2009, 15: 5060-5072. 10.1158/1078-0432.CCR-08-2245CrossRefPubMed

42.

Garofalo M, Romano G, Di Leva G, Nuovo G, Jeon YJ, Ngankeu A, Sun J, Lovat F, Alder H, Condorelli G, Engelman JA, Ono M, Rho JK, Cascione L, Volinia S, Nephew KP, Croce CM: EGFR and MET receptor tyrosine kinase-altered microRNA expression induces tumorigenesis and gefitinib resistance in lung cancers. Nat Med. 2012, 18: 74-82.

43.

Zhang KL, Han L, Chen LY, Shi ZD, Yang M, Ren Y, Chen LC, Zhang JX, Pu PY, Kang CS: Blockage of a miR-21/EGFR regulatory feedback loop augments anti-EGFR therapy in glioblastomas. Cancer Lett. 2014, 342: 139-149. 10.1016/j.canlet.2013.08.043CrossRefPubMed

Title: MicroRNA-566 activates EGFR signaling and its inhibition sensitizes glioblastoma cells to nimotuzumab
Authors: Kai-Liang Zhang
Xuan Zhou
Lei Han
Lu-Yue Chen
Ling-Chao Chen
Zhen-Dong Shi
Ming Yang
Yu Ren
Jing-Xuan Yang
Thomas S Frank
Chuan-Bao Zhang
Jun-Xia Zhang
Pei-Yu Pu
Jian-Ning Zhang
Tao Jiang
Eric J Wagner
Min Li
Chun-Sheng Kang
Publication date: 01-12-2014
Publisher: BioMed Central
Published in: Molecular Cancer / Issue 1/2014
Electronic ISSN: 1476-4598
DOI: https://doi.org/10.1186/1476-4598-13-63

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