High-throughput retroviral tagging to identify genes and signaling pathways cooperating with BRAF in thyroid carcinogenesis

癌基因单独激活通常不足以转化上皮细胞。对于突变型BRAF驱动的PTC，我们试图揭示赋予癌细胞恶性表型所需的其他遗传异常。为了这个目的，作为一个模型，我们进行了逆转录病毒插入突变的强力霉素诱导BRAFV600 E表达克隆系来自大鼠甲状腺PCCL 3细胞，并挑选克隆，不需要TSH的增长。通过PCR扩增和基因芯片分析，我们成功地鉴定出了几个可能的候选基因（Trpm 1，Nek 6等）。

Activation of oncogene alone is usually insufficient to transform epithelial cells. For PTCs driven by mutant BRAF we attempted to reveal the additional genetic abnormalities that are required to confer malignant phenotype on cancer cells. For this purpose, as a model, we performed retrovirus insertional mutagenesis in doxycycline-inducible BRAFV600E-expressing clonal lines derived from rat thyroid PCCL3 cells, and picked the clones that do not require TSH for growth. By PCR-based splinkerette procedure and microarray analysis we have successfully identified several possible candidate genes (Trpm1, Nek6, etc).