Department of Molecular Medicine, Atomic Bomb Disease Institute, Nagasaki University, Nagasaki, Japan.
Department of Medicine and Molecular Science, Graduate School of Medicine, Gunma University, Maebashi, Japan.
PLoS One. 2018 Aug 7;13(8):e0201365. doi: 10.1371/journal.pone.0201365. eCollection 2018.
The BRAFV600E mutation is the most prevalent driver mutation of sporadic papillary thyroid cancers (PTC). It was previously shown that prenatal or postnatal expression of BRAFV600E under elevated TSH levels induced thyroid cancers in several genetically engineered mouse models. In contrast, we found that postnatal expression of BRAFV600E under physiologic TSH levels failed to develop thyroid cancers in conditional transgenic Tg(LNL-BrafV600E) mice injected in the thyroid with adenovirus expressing Cre under control of the thyroglobulin promoter (Ad-TgP-Cre). In this study, we first demonstrated that BrafCA/+ mice carrying a Cre-activated allele of BrafV600E exhibited higher transformation efficiency than Tg(LNL-BrafV600E) mice when crossed with TPO-Cre mice. As a result, most BrafCA/+ mice injected with Ad-TgP-Cre developed thyroid cancers in 1 year. Histologic examination showed follicular or cribriform-like structures with positive TG and PAX staining and no colloid formation. Some tumors also had papillary structure component with lower TG expression. Concomitant PTEN haploinsufficiency in injected BrafCA/+;Ptenf/+ mice induced tumors predominantly exhibiting papillary structures and occasionally undifferentiated solid patterns with normal to low PAX expression and low to absent TG expression. Typical nuclear features of human PTC and extrathyroidal invasion were observed primarily in the latter mice. The percentages of pERK-, Ki67- and TUNEL-positive cells were all higher in the latter. In conclusion, we established novel thyroid cancer mouse models in which postnatal expression of BRAFV600E alone under physiologic TSH levels induces PTC. Simultaneous PTEN haploinsufficiency tends to promote tumor growth and de-differentiation.
BRAFV600E 突变是散发性甲状腺乳头状癌(PTC)最常见的驱动突变。先前的研究表明,在高 TSH 水平下,BRAFV600E 的产前或产后表达会在几种基因工程小鼠模型中诱导甲状腺癌。相比之下,我们发现,在生理 TSH 水平下,BRAFV600E 的产后表达未能在注射甲状腺球蛋白启动子控制下的腺病毒表达 Cre(Ad-TgP-Cre)的条件性转基因 Tg(LNL-BrafV600E)小鼠中发展为甲状腺癌。在这项研究中,我们首先证明,与 TPO-Cre 小鼠杂交时,携带 Cre 激活的 BrafV600E 等位基因的 BrafCA/+ 小鼠比 Tg(LNL-BrafV600E) 小鼠具有更高的转化效率。结果,大多数注射 Ad-TgP-Cre 的 BrafCA/+ 小鼠在 1 年内发展为甲状腺癌。组织学检查显示,具有阳性 TG 和 PAX 染色且无胶体形成的滤泡或筛状结构。一些肿瘤还具有具有较低 TG 表达的乳头状结构成分。在注射的 BrafCA/+;Ptenf/+ 小鼠中同时存在 PTEN 杂合性缺失会诱导主要表现为乳头状结构的肿瘤,偶尔也表现为未分化的实性模式,具有正常至低水平的 PAX 表达和低至无 TG 表达。在后一种小鼠中主要观察到人类 PTC 的典型核特征和甲状腺外浸润。在后一种小鼠中,pERK-、Ki67-和 TUNEL 阳性细胞的百分比均较高。总之,我们建立了新的甲状腺癌小鼠模型,其中 BRAFV600E 的单独产后表达在生理 TSH 水平下可诱导 PTC。同时存在的 PTEN 杂合性缺失倾向于促进肿瘤生长和去分化。