Acceleration of BRAF V600E -induced thyroid carcinogenesis by TGFβ signal deficiency in mice

Acceleration of BRAF V600E -induced thyroid carcinogenesis by TGFβ signal deficiency in mice:

Abstract

Purpose

Transforming growth factor-β (TGFβ) has pleiotropic actions, including both anti- and pro-tumorigenic abilities. We have previously shown no tumor development in the thyroid-specific TGFβ receptor type II knockout (Tgfβr2 KO) mice, indicating the insufficiency of defective TGFβ signal itself for thyroid cancer initiation. In the current study, we evaluated whether defective TGFβ signal accelerates BRAF^V600E-mediated thyroid carcinogenesis in our mouse model, in which intrathyroidal injection of adenovirus expressing Cre under thyroglobulin (TG) promoter (Ad-TgP-Cre) into thyroid lobes of conditional Braf^V600E knock-in mice (Braf^CA) induces thyroid cancers 12 months later.

Methods

Braf^CA/wt;Tgfbr2^{floxE2/floxE2} mice were generated by crossing Tgfbr2^{floxE2/floxE2} and Braf^CA mice, and Ad-TgP-Cre was injected into the left lobes of 4–6-week-old mice. Mice were sacrificed at 6 and 12 months, and the thyroid tissues were subjected to H&E and immune-histochemistry and -fluorecence.

Results

Thyroid tumors were observed in 8 of 10 mice at 6 months and 4 of 7 mice at 12 months. These tumors were judged to be malignant by H&E staining, because of the presence of papillary growth of atypical follicular cells, intranuclear cytoplasmic inclusions and so on. Immunohistochemical analyses using thyroid cancer tissues obtained at 6 months demonstrated variable levels of TG but steady levels of Paired Box-8 expression and higher Ki67 positivity. The degree of epithelial-to-mesenchymal transition could not be evaluated because normal thyroid tissues and thyroid cancers developed in Braf^CA and Braf^CA/wt;Tgfbr2^{floxE2/floxE2} mice were all E-cadherin⁺/vimentin⁻, that is, epithelial type.

Conclusion

In a mouse model, defective TGFβ signaling pathway accelerates BRAF^V600E-induced thyroid cancer development, which is occasionally accompanied by reduced TG expression implying dedifferentiation. The former finding is consistent with anti-tumorigenic ability of TGFβ in early tumorigenic process, but the latter is contradictory to generally accepted concept for TGFβ-induction of dedifferentiation.