Microrna-199a-5p Functions as a Tumor Suppressor via Suppressing Connective Tissue Growth Factor (CTGF) in Follicular Thyroid Carcinoma

Background The objective of this study was to explore the role of miR-199a-5p in the development of thyroid cancer, including its anti-proliferation effect and downstream signaling pathway. Material/Methods We conducted qRT-PCR analysis to detect the expressions of several microRNAs in 42 follicular thyroid carcinoma patients and 42 controls. We identified CTGF as target of miR-491, and viability and cell cycle status were determined in FTC-133 cells transfected with CTGF siRNA, miR-199a mimics, or inhibitors. Results We identified an underexpression of miR-199a-5p in follicular thyroid carcinoma tissue samples compared with controls. Then we confirmed CTGF as a target of miR-199a-5p thyroid cells by using informatics analysis and luciferase reporter assay. Additionally, we found that mRNA and protein expression levels of CTGF were both clearly higher in malignant tissues than in benign tissues. miR-199a-5p mimics and CTGF siRNA similarly downregulated the expression of CTGF, and reduced the viability of FTC-133 cells by arresting the cell cycle in G0 phase. Transfection of miR-199a-5p inhibitors increased the expression of CTGF and promoted the viability of the cells by increasing the fraction of cells in G2/M and S phases. Conclusions Our study proves that the CTGF gene is a target of miR-199a-5p, demonstrating the negatively related association between CTGF and miR-199a. These findings suggest that miR-199a-5p might be a novel therapeutic target in the treatment of follicular thyroid carcinoma.

[1]  Jian Li,et al.  MicroRNA-199a-3p is downregulated in gastric carcinomas and modulates cell proliferation. , 2013, Genetics and molecular research : GMR.

[2]  R. Aharonov,et al.  MicroRNAs accurately identify cancer tissue origin , 2008, Nature Biotechnology.

[3]  India,et al.  Cancer Incidence in Five Continents Vol. X , 2013 .

[4]  Feng Yang,et al.  Stromal expression of connective tissue growth factor promotes angiogenesis and prostate cancer tumorigenesis. , 2005, Cancer research.

[5]  P. Bork.  The modular architecture of a new family of growth regulators related to connective tissue growth factor , 1993, FEBS letters.

[6]  Abnormal anion exchange mechanism operates in the sweat glands of cystic fibrosis patients. , 1984, Pediatric research.

[7]  H. Tao,et al.  Up-regulated miR-199a-5p in gastric cancer functions as an oncogene and targets klotho , 2014, BMC Cancer.

[8]  V. Ambros.  The functions of animal microRNAs , 2004, Nature.

[9]  L. Hegedüs,et al.  Management of simple nodular goiter: current status and future perspectives. , 2003, Endocrine reviews.

[10]  Yuri E Nikiforov,et al.  RAS point mutations and PAX8-PPAR gamma rearrangement in thyroid tumors: evidence for distinct molecular pathways in thyroid follicular carcinoma. , 2003, The Journal of clinical endocrinology and metabolism.

[11]  B. Perbal.  CCN proteins: multifunctional signalling regulators , 2004, The Lancet.

[12]  X. Wan,et al.  MicroRNA‐199a targets CD44 to suppress the tumorigenicity and multidrug resistance of ovarian cancer‐initiating cells , 2012, The FEBS journal.

[13]  Thomas D. Schmittgen,et al.  miR-199a-3p targets CD44 and reduces proliferation of CD44 positive hepatocellular carcinoma cell lines. , 2010, Biochemical and biophysical research communications.

[14]  Yuri E. Nikiforov,et al.  Diagnostic pathology and molecular genetics of the thyroid , 2012 .

[15]  R. Seethala,et al.  Impact of mutational testing on the diagnosis and management of patients with cytologically indeterminate thyroid nodules: a prospective analysis of 1056 FNA samples. , 2011, The Journal of clinical endocrinology and metabolism.

[16]  S. Hwang,et al.  Altered MicroRNA Expression in Cervical Carcinomas , 2008, Clinical Cancer Research.

[17]  F. Pacini,et al.  Impact of proto-oncogene mutation detection in cytological specimens from thyroid nodules improves the diagnostic accuracy of cytology. , 2010, The Journal of clinical endocrinology and metabolism.

[18]  Dong Xie,et al.  Levels of Expression of CYR61 and CTGF Are Prognostic for Tumor Progression and Survival of Individuals with Gliomas , 2004, Clinical Cancer Research.

[19]  D. Xie,et al.  Connective Tissue Growth Factor Is Overexpressed in Esophageal Squamous Cell Carcinoma and Promotes Tumorigenicity through β-Catenin-T-cell Factor/Lef Signaling* , 2007, Journal of Biological Chemistry.

[20]  Jae Hoon Kim,et al.  MicroRNA Expression Profiles in Serous Ovarian Carcinoma , 2008, Clinical Cancer Research.

[21]  King-Jen Chang,et al.  Connective tissue growth factor confers drug resistance in breast cancer through concomitant up-regulation of Bcl-xL and cIAP1. , 2009, Cancer research.

[22]  Kazuhiro Yoshida,et al.  Relation between microRNA expression and progression and prognosis of gastric cancer: a microRNA expression analysis. , 2010, The Lancet. Oncology.

[23]  H. Inoue,et al.  Expression of connective tissue growth factor in cartilaginous tumors , 2000, Cancer.

[24]  M. Nikiforova,et al.  Molecular testing for mutations in improving the fine-needle aspiration diagnosis of thyroid nodules. , 2009, The Journal of clinical endocrinology and metabolism.

[25]  R. DeLellis.  Pathology and genetics of tumours of endocrine organs , 2004 .

[26]  C. Eng,et al.  A limited set of human MicroRNA is deregulated in follicular thyroid carcinoma. , 2006, The Journal of clinical endocrinology and metabolism.

[27]  H. Moch,et al.  MicroRNA expression array identifies novel diagnostic markers for conventional and oncocytic follicular thyroid carcinomas. , 2013, The Journal of clinical endocrinology and metabolism.

[28]  M. Sobrinho-Simões,et al.  Follicular thyroid carcinoma , 2011, Modern Pathology.

[29]  B. Reinhart,et al.  Prediction of Plant MicroRNA Targets , 2002, Cell.

[30]  Yu Liang,et al.  BMC Genomics , 2007 .

[31]  T. Fahey,et al.  MicroRNA analysis as a potential diagnostic tool for papillary thyroid carcinoma , 2008, Modern Pathology.

[32]  Yan Wang,et al.  Expression of Cyr61, CTGF, and WISP-1 Correlates with Clinical Features of Lung Cancer , 2007, PloS one.

[33]  K. Lyons,et al.  CCN family 2/connective tissue growth factor (CCN2/CTGF) regulates the expression of Vegf through Hif-1alpha expression in a chondrocytic cell line, HCS-2/8, under hypoxic condition. , 2009, Bone.

[34]  D. Bartel.  MicroRNAs Genomics, Biogenesis, Mechanism, and Function , 2004, Cell.

[35]  King-Jen Chang,et al.  Connective tissue growth factor inhibits metastasis and acts as an independent prognostic marker in colorectal cancer. , 2005, Gastroenterology.