MicroRNA‑9‑5p functions as a tumor suppressor in papillary thyroid cancer via targeting BRAF

Guo,Feng; Hou,Xinming; Sun,Qinghui

doi:10.3892/ol.2018.9423

MicroRNA‑9‑5p functions as a tumor suppressor in papillary thyroid cancer via targeting BRAF

Authors:
- Feng Guo
- Xinming Hou
- Qinghui Sun
View Affiliations

Affiliations: Department of Thyroid and Breast Surgery, The Second People's Hospital of Liaocheng, Liaocheng, Shandong 252600, P.R. China
Published online on: September 7, 2018 https://doi.org/10.3892/ol.2018.9423
Pages: 6815-6821

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

MicroRNAs (miRNAs/miRs) are widely studied as key regulators of gene expression and are involved in various diseases by affecting the miRNA‑mediated regulatory function. BRAF is an important oncogene in the regulation of cell proliferation and apoptosis. In the present study, reverse transcription‑quantitative polymerase chain reaction was used to determine the expression levels of miR‑9‑5p and BRAF mRNA in patients with papillary thyroid cancer (PTC). Western blotting was used to detect BRAF protein level. A luciferase assay was used to verify the miR‑9‑5p target site in BRAF. Cell Counting Kit‑8 and flow cytometry were used to assess cell proliferation, and apoptosis, respectively. In the present study, it was demonstrated that miR‑9‑5p is downregulated in malignant PTC. Using bioinformatics analysis, miR‑9‑5p was predicted to target the human BRAF 3'‑untranslated region (3'‑UTR). A dual‑luciferase assay demonstrated that miR‑9‑5p downregulated BRAF expression by directly targeting its 3'‑UTR. Mutations in the 3'‑UTR of BRAF completely abolished its interaction with miR‑9‑5p. Expression of exogenous miRNA that mimics miR‑9‑5p miRNA decreased BRAF protein and mRNA levels, while suppression of endogenous miR‑9‑5p resulted in an increase in BRAF protein, and mRNA levels. Furthermore, regulation of miR‑9‑5p was observed to suppress the viability of PTC cells by inducing apoptosis. Consistently, downregulation of miR‑9‑5p promoted proliferation of PTC cells by inhibiting the apoptosis of cells. In conclusion, the present study demonstrated that miR‑9‑5p may perform an important role in PTC prognosis and therapy.

View Figures

View References

1	Al-Brahim N and Asa SL: Papillary thyroid carcinoma: An overview. Arch Pathol Lab Med. 130:1057–1062. 2006.PubMed/NCBI
2	Segovia Gomez I, Gallowitsch HJ, Kresnik E, Kumnig G, Igerc I, Matschnig S, Stronegger WJ and Lind P: Descriptive epidemiology of thyroid carcinoma in Carinthia, Austria: 1984–2001. Histopathologic features and tumor classification of 734 cases under elevated general iodination of table salt since 1990: Population-based age-stratified analysis on thyroid carcinoma incidence. Thyroid. 14:277–286. 2004. View Article : Google Scholar : PubMed/NCBI
3	Akslen LA, Haldorsen T, Thoresen SO and Glattre E: Incidence pattern of thyroid cancer in Norway: Influence of birth cohort and time period. Int J Cancer. 53:183–187. 1993. View Article : Google Scholar : PubMed/NCBI
4	Colonna M, Grosclaude P, Remontet L, Schvartz C, Mace-Lesech J, Velten M, Guizard A, Tretarre B, et al: Incidence of thyroid cancer in adults recorded by French cancer registries (1978–1997). Eur J Cancer. 38:1762–1768. 2002. View Article : Google Scholar : PubMed/NCBI
5	Howe HL, Wingo PA, Thun MJ, Ries LA, Rosenberg HM, Feigal EG and Edwards BK: Annual report to the nation on the status of cancer (1973 through 1998), featuring cancers with recent increasing trends. J Natl Cancer Inst. 93:824–842. 2001. View Article : Google Scholar : PubMed/NCBI
6	Davies L and Welch HG: Increasing incidence of thyroid cancer in the United States, 1973–2002. JAMA. 295:2164–2167. 2006. View Article : Google Scholar : PubMed/NCBI
7	Churilla TM, Donnelly PE, Leatherman ER, Adonizio CS and Peters CA: Total mastectomy or breast conservation therapy? how radiation oncologist accessibility determines treatment choice and quality: A SEER Data-base analysis. Breast J. 21:473–480. 2015. View Article : Google Scholar : PubMed/NCBI
8	Schechter RB, Nagilla M, Joseph L, Reddy P, Khattri A, Watson S, Locati LD, Licitra L, Greco A, Pelosi G, et al: Genetic profiling of advanced radioactive iodine-resistant differentiated thyroid cancer and correlation with axitinib efficacy. Cancer Lett. 359:269–274. 2015. View Article : Google Scholar : PubMed/NCBI
9	Knauf JA, Ma X, Smith EP, Zhang L, Mitsutake N, Liao XH, Refetoff S, Nikiforov YE and Fagin JA: Targeted expression of BRAFV600E in thyroid cells of transgenic mice results in PTCs that undergo dedifferentiation. Cancer Res. 65:4238–4245. 2005. View Article : Google Scholar : PubMed/NCBI
10	Kundu A, Quirit JG, Khouri MG and Firestone GL: Inhibition of oncogenic BRAF activity by indole-3-carbinol disrupts microphthalmia-associated transcription factor expression and arrests melanoma cell proliferation. Mol Carcinog. 56:49–61. 2017. View Article : Google Scholar : PubMed/NCBI
11	Saugstad JA: MicroRNAs as effectors of brain function with roles in ischemia and injury, neuroprotection, and neurodegeneration. J Cereb Blood Flow Metab. 30:1564–1576. 2010. View Article : Google Scholar : PubMed/NCBI
12	Nalysnyk L, Cid-Ruzafa J, Rotella P and Esser D: Incidence and prevalence of idiopathic pulmonary fibrosis: Review of the literature. Eur Respir Rev. 21:355–361. 2012. View Article : Google Scholar : PubMed/NCBI
13	Sondermann A, Andreghetto FM, Moulatlet AC, da Silva Victor E, de Castro MG, Nunes FD, Brandão LG and Severino P: MiR-9 and miR-21 as prognostic biomarkers for recurrence in PTC. Clin Exp Metastasis. 32:521–530. 2015. View Article : Google Scholar : PubMed/NCBI
14	Yoon S, An YS, Lee SJ, So EY, Kim JH, Chung YS and Yoon JK: Relation between F-18 FDG uptake of PET/CT and BRAFV600E mutation in PTC. Medicine (Baltimore). 94:e20632015. View Article : Google Scholar : PubMed/NCBI
15	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–8. 2001. View Article : Google Scholar : PubMed/NCBI
16	Shen CH, Yuan P, Perez-Lorenzo R, Zhang Y, Lee SX, Ou Y, Asara JM, Cantley LC and Zheng B: Phosphorylation of BRAF by AMPK impairs BRAF-KSR1 association and cell proliferation. Mol Cell. 52:161–172. 2013. View Article : Google Scholar : PubMed/NCBI
17	Rothenberg SM, Daniels GH and Wirth LJ: Redifferentiation of iodine-refractory BRAF V600E-Mutant metastatic papillary thyroid cancer with dabrafenib-response. Clin Cancer Res. 21:5640–5641. 2015. View Article : Google Scholar : PubMed/NCBI
18	Noble PW, Barkauskas CE and Iang D: Pulmonary fibrosis: Patterns and perpetrators. J Clin Invest. 122:2756–2762. 2012. View Article : Google Scholar : PubMed/NCBI
19	Amara N, Goven D, Prost F, Muloway R, Crestani B and Boczkowski J: NOX4/NADPH oxidase expression is increased in pulmonary fibroblasts from patients with idiopathic pulmonary fibrosis and mediates TGFbeta1-induced fibroblast differentiation into myofibroblasts. Thorax. 65:733–738. 2010. View Article : Google Scholar : PubMed/NCBI
20	Udell CM, Rajakulendran T, Sicheri F and Therrien M: Mechanistic principles of RAF kinase signaling. Cell Mol Life Sci. 68:553–565. 2011. View Article : Google Scholar : PubMed/NCBI
21	Osborne JK, Zaganjor E and Cobb MH: Signal control through Raf: In sickness and in health. Cell Res. 22:14–22. 2012. View Article : Google Scholar : PubMed/NCBI
22	Roberts PJ and Der V: Targeting the Raf-MEK-ERK mitogen-activated protein kinase cascade for the treatment of cancer. Oncogene. 26:3291–3310. 2007. View Article : Google Scholar : PubMed/NCBI
23	Matallanas D, Birtwistle M, Romano D, Zebisch A, Rauch J, von Kriegsheim A and Kolch W: Raf Family Kinases: Old dogs have learned new tricks. Genes Cancer. 2:232–260. 2011. View Article : Google Scholar : PubMed/NCBI
24	Gray-Schopfer V, Wellbrock C and Marais R: Melanoma biology and new targeted therapy. Nature. 445:851–857. 2007. View Article : Google Scholar : PubMed/NCBI
25	American Thyroid Association (ATA) Guidelines Taskforce on Thyroid Nodules and Differentiated Thyroid Cancer, . Cooper DS, Doherty GM, Haugen BR, Kloos RT, Lee SL, Mandel SJ, Mazzaferri EL, McIver B, Pacini F, et al: Revised American Thyroid Association management guidelines for patients with thyroid nodules and differentiated thyroid cancer. Thyroid. 19:1167–1214. 2009. View Article : Google Scholar : PubMed/NCBI
26	Tuttle RM, Ball DW, Byrd D, Dilawari RA, Doherty GM, Duh QY, Ehya H, Farrar WB, Haddad RI, Kandeel F, et al: Thyroid carcinoma. J Natl Compr Canc Netw. 8:1228–1274. 2010. View Article : Google Scholar : PubMed/NCBI
27	Brown RL, de Souza JA and Cohen EE: Thyroid cancer: Burden of illness and management of disease. J Cancer. 2:193–199. 2011. View Article : Google Scholar : PubMed/NCBI
28	Cohen Y, Xing M, Mambo E, Guo Z, Wu G, Trink B, Beller U, Westra WH, Ladenson PW and Sidransky D: BRAF mutation in papillary thyroid carcinoma. J Natl Cancer Inst. 95:625–627. 2003. View Article : Google Scholar : PubMed/NCBI
29	Lin KL, Wang OC, Zhang XH, Dai XX, Hu XQ and Qu JM: The BRAF mutation is predictive of aggressive clinicopathological characteristics in papillary thyroid microcarcinoma. Ann Surg Oncol. 1:3294–3300. 2010. View Article : Google Scholar
30	Kimura ET, Nikiforova MN, Zhu Z, Knauf JA, Nikiforov YE and Fagin JA: High prevalence of BRAF mutations in thyroid cancer: Genetic evidence for constitutive activation of the RET/PTC-RAS-BRAF signaling pathway in papillary thyroid carcinoma. Cancer Res. 63:1454–1457. 2003.PubMed/NCBI
31	Soares P, Trovisco V, Rocha AS, Lima J, Castro P, Preto A, Máximo V, Botelho T, Seruca R and Sobrinho-Simões M: BRAF mutations and RET/PTC rearrangements are alternative events in the etiopathogenesis of PTC. Oncogene. 22:4578–4580. 2003. View Article : Google Scholar : PubMed/NCBI
32	Vanden Borre P, McFadden DG, Gunda V, Sadow PM, Varmeh S, Bernasconi M, Jacks T and Parangi S: The next generation of orthotopic thyroid cancer models: immunocompetent orthotopic mouse models of BRAF V600E-positive papillary and anaplastic thyroid carcinoma. Thyroid. 24:705–714. 2014. View Article : Google Scholar : PubMed/NCBI
33	Xing M, Alzahrani AS, Carson KA, Viola D, Elisei R, Bendlova B, Yip L, Mian C, Vianello F, Tuttle RM, et al: Association between BRAF V600E mutation and mortality in patients with papillary thyroid cancer. JAMA. 309:1493–1501. 2013. View Article : Google Scholar : PubMed/NCBI
34	Tang HC and Chen YC: Insight into molecular dynamics simulation of BRAF(V600E) and potent novel inhibitors for malignant melanoma. Int J Nanomedicine. 10:3131–3146. 2015.PubMed/NCBI
35	Eloy C, Santos J, Soares P and Sobrinho-Simões M: The preeminence of growth pattern and invasiveness and the limited influence of BRAF and RAS mutations in the occurrence of papillary thyroid carcinoma lymph node metastases. Virchows Arch. 459:265–276. 2011. View Article : Google Scholar : PubMed/NCBI
36	Cheng S, Serra S, Mercado M, Ezzat S and Asa SL: A high-throughput proteomic approach provides distinct signatures for thyroid cancer behavior. Clin Cancer Res. 17:2385–2394. 2011. View Article : Google Scholar : PubMed/NCBI