J Korean Surg Soc.  2013 Jan;84(1):9-17.

Osteopontin expression in papillary thyroid carcinoma and its relationship with the BRAF mutation and tumor characteristics

• Kang KH

Abstract

PURPOSE
We evaluated the relationship between the degree of osteopontin (OPN) expression in papillary thyroid carcinoma (PTC) specimens and the presence of the BRAF mutation and clinicopathologic variables.
METHODS
Fifty-six snap-frozen thyroid tumor samples, including those of 49 PTCs, four follicular adenomas, two follicular carcinomas, and one Hurthle cell adenoma, were studied. We performed reverse transcription-polymerase chain reaction (RT-PCR) to assess the OPN expression levels. We also tested the BRAF codon 599 mutations using RT-PCR with the direct sequencing method. All of the tumors were microscopically reexamined by a pathologist with a special interest in thyroid neoplasia.
RESULTS
OPN mRNA was significantly overexpressed in the PTC samples compared with other thyroid tumors (P = 0.011). In PTCs, the OPN expression level was higher in the BRAF mutation group than in the wild-type group (P = 0.041). Among the clinicopathologic variables, nonfollicular variant histologic subtypes (P = 0.002) and the presence of lymph node metastases (P = 0.042) were correlated with elevated level of OPN expression.
CONCLUSION
OPN might play a crucial role in tumorigenesis and the progression of PTC.

Keyword

Osteoponin; Papillary thyroid cancer

MeSH Terms

Adenoma
Carcinoma
Cell Transformation, Neoplastic
Codon
Factor IX
Lymph Nodes
Neoplasm Metastasis
Osteopontin
RNA, Messenger
Thyroid Gland
Thyroid Neoplasms
Carcinoma
Codon
Factor IX
Osteopontin
RNA, Messenger
Thyroid Neoplasms

Figure

• 

  Fig. 1 Osteopontin (OPN) expression according to histologic type. Each product of reverse transcription-polymerase chain reaction of OPN mRNA was loaded onto a 1.5% agarose gel that was stained with ethidium bromide (0.5 µg/mL). MyImager 1000 Image Analysis System was used for the quantification of each band. OPN mRNA expression of each sample was recorded as the fold increase with respect to the expression of the housekeeping GAPDH gene. CP, classic papillary type of papillary thyroid carcinoma; FV, follicular variant of papillary thyroid carcinoma; FA, follicular adenoma.

• 

  Fig. 2 Sequence chromatograms of BRAF exon 15 wild-type and mutant type of PTC BRAF mutation-positive sample (mutant type) shows the heterogeneous thymine-to-adenine transversion at nucleotide position 1796, which results in a valine-to-glutamate substitution at residue 599. All of the detected BRAF mutations in this study were these BRAFV599E mutations.

Reference

1. DeLellis RA, Williams ED. DeLellis LA, Lloyd RV, Heitz PU, Eng C, editors. Thyroid and parathyroid tumors. Pathology and genetics of tumours of endocrine organs. 2004. Lyon: IARC Press;51–56.

2. Pelizzo MR, Boschin IM, Toniato A, Pagetta C, Piotto A, Bernante P, et al. Natural history, diagnosis, treatment and outcome of papillary thyroid microcarcinoma (PTMC): a mono-institutional 12-year experience. Nucl Med Commun. 2004. 25:547–552.

3. Takahashi M, Ritz J, Cooper GM. Activation of a novel human transforming gene, ret, by DNA rearrangement. Cell. 1985. 42:581–588.

4. Airaksinen MS, Saarma M. The GDNF family: signalling, biological functions and therapeutic value. Nat Rev Neurosci. 2002. 3:383–394.

5. Grieco M, Santoro M, Berlingieri MT, Melillo RM, Donghi R, Bongarzone I, et al. PTC is a novel rearranged form of the ret proto-oncogene and is frequently detected in vivo in human thyroid papillary carcinomas. Cell. 1990. 60:557–563.

6. Tallini G, Asa SL. RET oncogene activation in papillary thyroid carcinoma. Adv Anat Pathol. 2001. 8:345–354.

7. Kondo T, Ezzat S, Asa SL. Pathogenetic mechanisms in thyroid follicular-cell neoplasia. Nat Rev Cancer. 2006. 6:292–306.

8. Kimura ET, Nikiforova MN, Zhu Z, Knauf JA, Nikiforov YE, 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. 2003. 63:1454–1457.

9. Cohen Y, Xing M, Mambo E, Guo Z, Wu G, Trink B, et al. BRAF mutation in papillary thyroid carcinoma. J Natl Cancer Inst. 2003. 95:625–627.

10. Davies H, Bignell GR, Cox C, Stephens P, Edkins S, Clegg S, et al. Mutations of the BRAF gene in human cancer. Nature. 2002. 417:949–954.

11. Zhu Z, Gandhi M, Nikiforova MN, Fischer AH, Nikiforov YE. Molecular profile and clinical-pathologic features of the follicular variant of papillary thyroid carcinoma: an unusually high prevalence of ras mutations. Am J Clin Pathol. 2003. 120:71–77.

12. Weber GF. The metastasis gene osteopontin: a candidate target for cancer therapy. Biochim Biophys Acta. 2001. 1552:61–85.

13. Rudland PS, Platt-Higgins A, El-Tanani M, De Silva, Barraclough R, Winstanley JH, et al. Prognostic significance of the metastasis-associated protein osteopontin in human breast cancer. Cancer Res. 2002. 62:3417–3427.

14. Guarino V, Faviana P, Salvatore G, Castellone MD, Cirafici AM, De Falco V, et al. Osteopontin is overexpressed in human papillary thyroid carcinomas and enhances thyroid carcinoma cell invasiveness. J Clin Endocrinol Metab. 2005. 90:5270–5278.

15. Livolsi VA. Surgical pathology of the thyroid. Philadelphia: Saunders;1990.

16. Rangaswami H, Bulbule A, Kundu GC. Osteopontin: role in cell signaling and cancer progression. Trends Cell Biol. 2006. 16:79–87.

17. Lin YH, Yang-Yen HF. The osteopontin-CD44 survival signal involves activation of the phosphatidylinositol 3-kinase/Akt signaling pathway. J Biol Chem. 2001. 276:46024–46030.

18. Das R, Mahabeleshwar GH, Kundu GC. Osteopontin stimulates cell motility and nuclear factor kappaB-mediated secretion of urokinase type plasminogen activator through phosphatidylinositol 3-kinase/Akt signaling pathways in breast cancer cells. J Biol Chem. 2003. 278:28593–28606.

19. Zheng DQ, Woodard AS, Tallini G, Languino LR. Substrate specificity of alpha(v)beta(3) integrin-mediated cell migration and phosphatidylinositol 3-kinase/AKT pathway activation. J Biol Chem. 2000. 275:24565–24574.

20. Nicholson KM, Anderson NG. The protein kinase B/Akt signalling pathway in human malignancy. Cell Signal. 2002. 14:381–395.

21. Castellone MD, Celetti A, Guarino V, Cirafici AM, Basolo F, Giannini R, et al. Autocrine stimulation by osteopontin plays a pivotal role in the expression of the mitogenic and invasive phenotype of RET/PTC-transformed thyroid cells. Oncogene. 2004. 23:2188–2196.

22. Gasparre G, Porcelli AM, Bonora E, Pennisi LF, Toller M, Iommarini L, et al. Disruptive mitochondrial DNA mutations in complex I subunits are markers of oncocytic phenotype in thyroid tumors. Proc Natl Acad Sci U S A. 2007. 104:9001–9006.

23. Giordano TJ, Kuick R, Thomas DG, Misek DE, Vinco M, Sanders D, et al. Molecular classification of papillary thyroid carcinoma: distinct BRAF, RAS, and RET/PTC mutation-specific gene expression profiles discovered by DNA microarray analysis. Oncogene. 2005. 24:6646–6656.

24. Tielens ET, Sherman SI, Hruban RH, Ladenson PW. Follicular variant of papillary thyroid carcinoma. A clinicopathologic study. Cancer. 1994. 73:424–431.

25. Jain M, Khan A, Patwardhan N, Reale F, Safran M. Follicular variant of papillary thyroid carcinoma: a comparative study of histopathologic features and cytology results in 141 patients. Endocr Pract. 2001. 7:79–84.

26. Puxeddu E, Moretti S, Elisei R, Romei C, Pascucci R, Martinelli M, et al. BRAF(V599E) mutation is the leading genetic event in adult sporadic papillary thyroid carcinomas. J Clin Endocrinol Metab. 2004. 89:2414–2420.

27. Kim KH, Kang DW, Kim SH, Seong IO, Kang DY. Mutations of the BRAF gene in papillary thyroid carcinoma in a Korean population. Yonsei Med J. 2004. 45:818–821.

28. Nikiforova MN, Kimura ET, Gandhi M, Biddinger PW, Knauf JA, Basolo F, et al. BRAF mutations in thyroid tumors are restricted to papillary carcinomas and anaplastic or poorly differentiated carcinomas arising from papillary carcinomas. J Clin Endocrinol Metab. 2003. 88:5399–5404.

29. Xing M, Westra WH, Tufano RP, Cohen Y, Rosenbaum E, Rhoden KJ, et al. BRAF mutation predicts a poorer clinical prognosis for papillary thyroid cancer. J Clin Endocrinol Metab. 2005. 90:6373–6379.