Abstract
The hybridisation of an Affymetrix HG_U95Av2 oligonucleotide array with RNAs extracted from six human thyroid carcinoma cell lines and a normal human thyroid primary cell culture led us to the identification of the UbcH10 gene that was upregulated by 150-fold in all of the carcinoma cell lines in comparison to the primary culture cells of human normal thyroid origin. Immunohistochemical studies performed on paraffin-embedded tissue sections showed abundant UbcH10 levels in thyroid anaplastic carcinoma samples, whereas no detectable UbcH10 expression was observed in normal thyroid tissues, in adenomas and goiters. Papillary and follicular carcinomas were only weakly positive. These results were further confirmed by RT–PCR and Western blot analyses. The block of UbcH10 protein synthesis induced by RNA interference significantly reduced the growth rate of thyroid carcinoma cell lines. Taken together, these results would indicate that UbcH10 overexpression is involved in thyroid cell proliferation, and may represent a marker of thyroid anaplastic carcinomas.
Highlights
The involvement of several oncogenes has been demonstrated in papillary thyroid carcinomas
To identify the genes regulated in the process of thyroid carcinogenesis, we analysed a microarray with RNAs extracted from normal human thyroid primary cell culture (NTPC), and six human thyroid carcinoma cell lines of different histotype
To search for candidate genes involved in the neoplastic transformation of thyroid gland, RNAs extracted from normal human thyroid primary cells and six human thyroid carcinoma cell lines of different origin (WRO cell line from a follicular carcinoma, TPC-1 and FB-2 cell lines, both deriving from papillary thyroid cancers, NPA cell line, which derives from a poorly differentiated papillary carcinoma, ARO and FRO cell lines originating from anaplastic carcinomas) were hybridised to U95Av2 Affymetrix oligonucleotide arrays
Summary
The involvement of several oncogenes has been demonstrated in papillary thyroid carcinomas. To identify the genes regulated in the process of thyroid carcinogenesis, we analysed a microarray with RNAs extracted from normal human thyroid primary cell culture (NTPC), and six human thyroid carcinoma cell lines of different histotype (one from a follicular carcinoma, three derived from papillary carcinomas and two from anaplastic carcinomas). The UbcH10 gene belongs to the E2 gene family and codes for a protein of 19.6 kDa that is involved in the ubiquitindependent proteolysis. In this pathway, ubiquitin-conjugating enzyme (E2), together with ubiquitin ligase (E3), transfers ubiquitin to specific substrate proteins (Hershko and Ciechanover, 1998; Joazeiro and Weissman, 2000). The block of UbcH10 protein synthesis by RNA interference inhibited the growth of two thyroid carcinoma cell lines
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