MicroRNA-146a inhibits epithelial mesenchymal transition in non-small cell lung cancer by targeting insulin receptor substrate 2

Park,Dong Ho; Jeon,Hyo  Sung; Lee,Soo Young; Choi,Yi  Young; Lee,Hae  Woo; Yoon,Seongkyu; Lee,Jae  Chel; Yoon,Yoo  Sang; Kim,Dae  Sung; Na,Moon  Jun; Kwon,Sun  Jung; Kim,Dong Sun; Kang,Jaeku; Park,Jae Yong; Son,Ji  Woong

doi:10.3892/ijo.2015.3111

MicroRNA-146a inhibits epithelial mesenchymal transition in non-small cell lung cancer by targeting insulin receptor substrate 2

Authors:
- Dong Ho Park
- Hyo Sung Jeon
- Soo Young Lee
- Yi Young Choi
- Hae Woo Lee
- Seongkyu Yoon
- Jae Chel Lee
- Yoo Sang Yoon
- Dae Sung Kim
- Moon Jun Na
- Sun Jung Kwon
- Dong Sun Kim
- Jaeku Kang
- Jae Yong Park
- Ji Woong Son
View Affiliations

Affiliations: Department of Anesthesiology and Pain Medicine, Konyang University Hospital, Daejeon, Republic of Korea, The Molecular Diagnostics and Imaging Research Institute, School of Medicine, Kyungpook National University, Daegu, Republic of Korea, Myunggok Research Institute for Medical Science, Konyang University, Daejeon, Republic of Korea, Department of Biochemistry, School of Medicine, Kyungpook National University, Daegu, Republic of Korea, Clinical Drug Manufacturing Center, Daegu-Gyeongbuk Medical Innovation Foundation, Daegu, Republic of Korea, Department of Chemical Engineering, University of Massachusetts Lowell, MA, USA, Department of Oncology, Asan Medical Center, University of Ulsan, College of Medicine, Seoul, Republic of Korea, Department of Thoracic Surgery, Konyang University Hospital, Daejeon, Republic of Korea, Department of Internal Medicine, Konyang University Hospital, Daejeon, Republic of Korea, Department of Anatomy, School of Medicine, Kyungpook National University, Daegu, Republic of Korea, Department of Pharmacology, College of Medicine, Konyang University, Daejeon, Republic of Korea, Lung Cancer Center, Kyungpook National University Medical Center, Daegu, Republic of Korea
Published online on: August 4, 2015 https://doi.org/10.3892/ijo.2015.3111
Pages: 1545-1553

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

During cancer progression, some tumor cells show changes in their plasticity by morphological and phenotypical conversions, as an expression of mesenchymal markers and loss of epithelial markers, collectively referred to as epithelial-mesenchymal transition (EMT). EMT has been increasingly recognized as a critical phenomenon in lung cancer progression. The goal of this study was to identify microRNAs involved in lung cancer progression. A microarray and qRT-PCR were performed to investigate the miRNA expression profiles in mesenchymal-like lung cancer cells. The role of miR‑146a in lung cancer progression was measured by invasion and migration assays in vitro. Bioinformatics and luciferase report assays were used to identify the target of miR‑146a. The expression of miR‑146a was reduced in mesenchymal-like lung cancer cell lines. The overexpression of miR‑146a induced a marked reduction of the mesenchymal marker and increase the epithelial marker in lung cancer cell lines. Moreover, the overexpression of miR‑146a suppressed lung cancer cell migration and invasion. Co-treatment with miR‑146a and gefitinib treatment showed a significant reduction of invasion in the resistant lung cancer cells induced by EMT. The expression of miR‑146a was downregulated in advanced lung cancer tissues. Insulin receptor substrate 2 (IRS2), an adaptor protein that modulates normal growth, metabolism, survival, and differentiation, was identified as a target of miR‑146a. miR‑146a regulated the expression of IRS2 at the mRNA and protein levels. These data demonstrate for the first time that miR‑146a suppresses lung cancer progression by repressing IRS2 expression. This provides new insight into the post-transcriptional regulation of lung cancer progression by miRNAs, a potential approach for the treatment of lung cancer.

View Figures

View References

1	Jemal A, Siegel R, Xu J and Ward E: Cancer statistics, 2010. CA Cancer J Clin. 60:277–300. 2010. View Article : Google Scholar : PubMed/NCBI
2	Yang P: Epidemiology of lung cancer prognosis: Quantity and quality of life. Cancer Epidemiology. Verma M: Humana Press; pp. 469–486. 2009, View Article : Google Scholar
3	Detterbeck FC: Lobectomy versus limited resection in T1N0 lung cancer. Ann Thorac Surg. 96:742–744. 2013. View Article : Google Scholar : PubMed/NCBI
4	van Zijl F, Krupitza G and Mikulits W: Initial steps of metastasis: Cell invasion and endothelial transmigration. Mutat Res. 728:23–34. 2011. View Article : Google Scholar : PubMed/NCBI
5	Shih J-Y and Yang P-C: The EMT regulator slug and lung carcinogenesis. Carcinogenesis. 32:1299–1304. 2011. View Article : Google Scholar : PubMed/NCBI
6	Sato M, Shames DS and Hasegawa Y: Emerging evidence of epithelial-to-mesenchymal transition in lung carcinogenesis. Respirology. 17:1048–1059. 2012. View Article : Google Scholar : PubMed/NCBI
7	Lin L and Bivona TG: Mechanisms of resistance to epidermal growth factor receptor inhibitors and novel therapeutic strategies to overcome resistance in NSCLC patients. Chemother Res Pract. 2012:8172972012.PubMed/NCBI
8	Jeon H-S, Lee YH, Lee SY, Jang JA, Choi YY, Yoo SS, Lee WK, Choi JE, Son JW, Kang YM, et al: A common polymorphism in pre-microRNA-146a is associated with lung cancer risk in a Korean population. Gene. 534:66–71. 2014. View Article : Google Scholar
9	Dearth RK, Cui X, Kim H-J, Hadsell DL and Lee AV: Oncogenic transformation by the signaling adaptor proteins insulin receptor substrate (IRS)-1 and IRS-2. Cell Cycle. 6:705–713. 2007. View Article : Google Scholar : PubMed/NCBI
10	Thomson S, Petti F, Sujka-Kwok I, Epstein D and Haley JD: Kinase switching in mesenchymal-like non-small cell lung cancer lines contributes to EGFR inhibitor resistance through pathway redundancy. Clin Exp Metastasis. 25:843–854. 2008. View Article : Google Scholar : PubMed/NCBI
11	Chung J-H, Rho JK, Xu X, Lee JS, Yoon HI, Lee CT, Choi YJ, Kim HR, Kim CH and Lee JC: Clinical and molecular evidences of epithelial to mesenchymal transition in acquired resistance to EGFR-TKIs. Lung Cancer. 73:176–182. 2011. View Article : Google Scholar
12	Chen J, Han Q and Pei D: EMT and MET as paradigms for cell fate switching. J Mol Cell Biol. 4:66–69. 2012. View Article : Google Scholar
13	Hay ED: The mesenchymal cell, its role in the embryo, and the remarkable signaling mechanisms that create it. Dev Dyn. 233:706–720. 2005. View Article : Google Scholar : PubMed/NCBI
14	Lee JM, Dedhar S, Kalluri R and Thompson EW: The epithelial-mesenchymal transition: New insights in signaling, development, and disease. J Cell Biol. 172:973–981. 2006. View Article : Google Scholar : PubMed/NCBI
15	Thiery JP and Sleeman JP: Complex networks orchestrate epithelial-mesenchymal transitions. Nat Rev Mol Cell Biol. 7:131–142. 2006. View Article : Google Scholar : PubMed/NCBI
16	Denlinger CE, Ikonomidis JS, Reed CE and Spinale FG: Epithelial to mesenchymal transition: The doorway to metastasis in human lung cancers. J Thorac Cardiovasc Surg. 140:505–513. 2010. View Article : Google Scholar : PubMed/NCBI
17	Thiery JP: Epithelial-mesenchymal transitions in tumour progression. Nat Rev Cancer. 2:442–454. 2002. View Article : Google Scholar : PubMed/NCBI
18	Nakata S, Sugio K, Uramoto H, Oyama T, Hanagiri T, Morita M and Yasumoto K: The methylation status and protein expression of CDH1, p16(INK4A), and fragile histidine triad in nonsmall cell lung carcinoma: Epigenetic silencing, clinical features, and prognostic significance. Cancer. 106:2190–2199. 2006. View Article : Google Scholar : PubMed/NCBI
19	Liu D, Huang C, Kameyama K, Hayashi E, Yamauchi A, Kobayashi S and Yokomise H: E-cadherin expression associated with differentiation and prognosis in patients with non-small cell lung cancer. Ann Thorac Surg. 71:949–954; discussion 954–945. 2001. View Article : Google Scholar : PubMed/NCBI
20	Hung J-J, Yang M-H, Hsu H-S, Hsu W-H, Liu J-S and Wu K-J: Prognostic significance of hypoxia-inducible factor-1α, TWIST1 and Snail expression in resectable non-small cell lung cancer. Thorax. 64:1082–1089. 2009. View Article : Google Scholar : PubMed/NCBI
21	Tischler V, Pfeifer M, Hausladen S, Schirmer U, Bonde AK, Kristiansen G, Sos ML, Weder W, Moch H, Altevogt P, et al: L1CAM protein expression is associated with poor prognosis in non-small cell lung cancer. Mol Cancer. 10:1272011. View Article : Google Scholar : PubMed/NCBI
22	Zhuo WL, Wang Y, Zhuo XL, Zhang YS and Chen ZT: Short interfering RNA directed against TWIST, a novel zinc finger transcription factor, increases A549 cell sensitivity to cisplatin via MAPK/mitochondrial pathway. Biochem Biophys Res Commun. 369:1098–1102. 2008. View Article : Google Scholar : PubMed/NCBI
23	Rho JK, Choi YJ, Lee JK, Ryoo BY, Na II, Yang SH, Kim CH and Lee JC: Epithelial to mesenchymal transition derived from repeated exposure to gefitinib determines the sensitivity to EGFR inhibitors in A549, a non-small cell lung cancer cell line. Lung Cancer. 63:219–226. 2009. View Article : Google Scholar
24	Friedman RC, Farh KK-H, Burge CB and Bartel DP: Most mammalian mRNAs are conserved targets of microRNAs. Genome Res. 19:92–105. 2009. View Article : Google Scholar :
25	Ceppi P and Peter ME: MicroRNAs regulate both epithelial-to-mesenchymal transition and cancer stem cells. Oncogene. 33:269–278. 2014. View Article : Google Scholar
26	Park S-M, Gaur AB, Lengyel E and Peter ME: The miR-200 family determines the epithelial phenotype of cancer cells by targeting the E-cadherin repressors ZEB1 and ZEB2. Genes Dev. 22:894–907. 2008. View Article : Google Scholar : PubMed/NCBI
27	Gregory PA, Bert AG, Paterson EL, Barry SC, Tsykin A, Farshid G, Vadas MA, Khew-Goodall Y and Goodall GJ: The miR-200 family and miR-205 regulate epithelial to mesenchymal transition by targeting ZEB1 and SIP1. Nat Cell Biol. 10:593–601. 2008. View Article : Google Scholar : PubMed/NCBI
28	Ceppi P, Mudduluru G, Kumarswamy R, Rapa I, Scagliotti GV, Papotti M and Allgayer H: Loss of miR-200c expression induces an aggressive, invasive, and chemoresistant phenotype in non-small cell lung cancer. Mol Cancer Res. 8:1207–1216. 2010. View Article : Google Scholar : PubMed/NCBI
29	Yu T, Li J, Yan M, Liu L, Lin H, Zhao F, Sun L, Zhang Y, Cui Y, Zhang F, et al: MicroRNA-193a-3p and -5p suppress the metastasis of human non-small-cell lung cancer by downregulating the ERBB4/PIK3R3/mTOR/S6K2 signaling pathway. Oncogene. 34:413–423. 2015. View Article : Google Scholar
30	Moustakas A and Heldin C-H: Signaling networks guiding epithelial-mesenchymal transitions during embryogenesis and cancer progression. Cancer Sci. 98:1512–1520. 2007. View Article : Google Scholar : PubMed/NCBI
31	Zhou L, Zhao X, Han Y, Lu Y, Shang Y, Liu C, Li T, Jin Z, Fan D and Wu K: Regulation of UHRF1 by miR-146a/b modulates gastric cancer invasion and metastasis. FASEB J. 27:4929–4939. 2013. View Article : Google Scholar : PubMed/NCBI
32	Karakatsanis A, Papaconstantinou I, Gazouli M, Lyberopoulou A, Polymeneas G and Voros D: Expression of microRNAs, miR-21, miR-31, miR-122, miR-145, miR-146a, miR-200c, miR-221, miR-222, and miR-223 in patients with hepatocellular carcinoma or intrahepatic cholangiocarcinoma and its prognostic significance. Mol Carcinog. 52:297–303. 2013. View Article : Google Scholar
33	Xu B, Wang N, Wang X, Tong N, Shao N, Tao J, Li P, Niu X, Feng N, Zhang L, et al: MiR-146a suppresses tumor growth and progression by targeting EGFR pathway and in a p-ERK-dependent manner in castration-resistant prostate cancer. Prostate. 72:1171–1178. 2012. View Article : Google Scholar
34	Chen G, Umelo IA, Lv S, Teugels E, Fostier K, Kronenberger P, Dewaele A, Sadones J, Geers C and De Grève J: miR-146a inhibits cell growth, cell migration and induces apoptosis in non-small cell lung cancer cells. PLoS One. 8:e603172013. View Article : Google Scholar : PubMed/NCBI
35	Carew RM, Browne MB, Hickey FB and Brazil DP: Insulin receptor substrate 2 and FoxO3a signalling are involved in E-cadherin expression and transforming growth factor-β1-induced repression in kidney epithelial cells. FEBS J. 278:3370–3380. 2011. View Article : Google Scholar : PubMed/NCBI
36	Geng Y, Ju Y, Ren F, Qiu Y, Tomita Y, Tomoeda M, Kishida M, Wang Y, Jin L, Su F, et al: Insulin receptor substrate 1/2 (IRS1/2) regulates Wnt/β-catenin signaling through blocking autophagic degradation of dishevelled2. J Biol Chem. 289:11230–11241. 2014. View Article : Google Scholar : PubMed/NCBI
37	Misra A, Pandey C, Sze SK and Thanabalu T: Hypoxia activated EGFR signaling induces epithelial to mesenchymal transition (EMT). PLoS One. 7:e497662012. View Article : Google Scholar : PubMed/NCBI
38	Cui X, Kim H-J, Kuiatse I, Kim H, Brown PH and Lee AV: Epidermal growth factor induces insulin receptor substrate-2 in breast cancer cells via c-Jun NH(2)-terminal kinase/activator protein-1 signaling to regulate cell migration. Cancer Res. 66:5304–5313. 2006. View Article : Google Scholar : PubMed/NCBI
39	Dearth RK, Cui X, Kim H-J, Kuiatse I, Lawrence NA, Zhang X, Divisova J, Britton OL, Mohsin S, Allred DC, et al: Mammary tumorigenesis and metastasis caused by overexpression of insulin receptor substrate 1 (IRS-1) or IRS-2. Mol Cell Biol. 26:9302–9314. 2006. View Article : Google Scholar : PubMed/NCBI
40	Chan BT and Lee AV: Insulin receptor substrates (IRSs) and breast tumorigenesis. J Mammary Gland Biol Neoplasia. 13:415–422. 2008. View Article : Google Scholar : PubMed/NCBI
41	Reuveni H, Flashner-Abramson E, Steiner L, Makedonski K, Song R, Shir A, Herlyn M, Bar-Eli M and Levitzki A: Therapeutic destruction of insulin receptor substrates for cancer treatment. Cancer Res. 73:4383–4394. 2013. View Article : Google Scholar : PubMed/NCBI