Growth factor production by human thyroid carcinoma cells: abundant expression of a platelet-derived growth factor-B-like protein by a human papillary carcinoma cell line

Abstract

As papillary thyroid carcinoma cells grow surrounding finger-like structures of stromal tissue, we postulated they may secrete a growth factor(s) for mesenchymal cells and that these would be distinct from any mitogenic factors elaborated by follicular carcinomas. Conditioned medium from both the human papillary carcinoma cell line NPA and the follicular carcinoma cell line WRO evoked a 20- to 30-fold increase in [3H]thymidine incorporation into NIH3T3 cell DNA. NPA cell growth factor activity largely eluted with 0.5 mol/L NaCl from a heparin-Sepharose column. NPA-conditioned medium competed in a platelet-derived growth factor-B (PDGF-B) RRA, and the mitogenic activity was partially blocked by an anti-PDGF-BB antibody. An immunoprecipitated PDGF-B-like protein from NPA cells was about 17 kilodaltons in a reducing gel, but, in contrast to wild-type PDGF-BB, did not change its electrophoretic mobility in an unreduced sodium dodecyl sulfate-polyacrylamide gel electrophoresis. NPA cells expressed an abundant 1.4-kilobase RNA that hybridized to probes for the 5'-untranslated and amino-terminal domains of PDGF-B and was distinct from the 4.2-kilobase wild-type PDGF-B chain transcript. There were no structural changes in the PDGF-B gene, as determined by cytogenetic analysis and restriction mapping. However, the PDGF-B gene in the NPA cells was hypomethylated compared to that in normal thyroid tissue or WRO cells. In contrast, the mitogenic activity of WRO cells bound to heparin with high affinity and was blocked by a basic fibroblast growth factor (bFGF) antibody. WRO cells contained abundant bFGF mRNA. Both cell lines abundantly expressed transforming growth factor-beta mRNA. Thus, NPA and WRO cells express powerful, yet distinct, mesenchymal cell growth factors. Whereas WRO cells express abundant bFGF, NPA cells produce a novel PDGF-B-like protein, which may correspond to a mutated form of PDGF-B-chain.