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

单独激活癌基因通常不足以转化上皮细胞。对于突变BRAF驱动的ptc，我们试图揭示赋予癌细胞恶性表型所需的额外遗传异常。为此，作为模型，我们对大鼠甲状腺PCCL3细胞衍生的强力霉素诱导的表达brafv600e的无性系进行了逆转录病毒插入突变，并挑选了不需要TSH生长的无性系。通过基于pcr的splinkerette程序和微阵列分析，我们成功地鉴定了几个可能的候选基因（Trpm1， Nek6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).