Long-term establishment, characterization and manipulation of cell lines from mouse basal cell carcinoma tumors
Long-term establishment, characterization and manipulation of cell lines from mouse basal cell carcinoma tumors
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DOI：
10.1111/j.1600-0625.2006.00465.x
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发表时间：
2006-09-01
影响因子：
3.6
通讯作者：
Epstein, Ervin H., Jr.
中科院分区：
文献类型：
--
作者：
So, Po-Lin;Langston, Alexander W.;Epstein, Ervin H., Jr.
关于从皮肤基底细胞癌 (BCC) 肿瘤中建立细胞的成功长期培养的报道很少。在这里，我们描述了使我们能够建立三种长期培养的 BCC 细胞的技术，这些细胞是从受辐射的 Patched 1 (Ptch1)(+/-) 小鼠中产生的 BCC 肿瘤中分离出来的。所有三种细胞系都显示出与 BCC 肿瘤相似的细胞形态，并且可以繁殖至少 20 代。此外，与 BCC 肿瘤类似，所有细胞系都丢失了野生型 Ptch1 等位基因，表达 BCC 分子标记，并对环杷明（一种 Hedgehog 信号传导的小分子抑制剂）有类似的反应。最后，我们描述了一种有效的电穿孔技术，用于将 DNA 转染到 BCC 细胞系中，并表明它们已激活 Hedgehog 信号活性，尽管其水平低于体内小鼠 BCC 的水平。这些数据表明该细胞系是真正的BCC细胞长期培养物，并且可以使用改进的电穿孔技术以相对高的转染效率将DNA质粒引入BCC细胞系中。
There have been few reports of successful long-term culture of cells established from cutaneous basal cell carcinoma (BCC) tumors. Here, we describe techniques that have enabled us to establish three long-term cultures of BCC cells isolated from BCC tumors that arose in irradiated Patched 1 (Ptch1)(+/-) mice. All three cell lines showed cellular morphology similar to that of BCC tumors and could be propagated for at least 20 passages. In addition, similar to BCC tumors, all cell lines had lost the wildtype Ptch1 allele, expressed BCC molecular markers, and responded similarly to cyclopamine, a small molecule inhibitor of Hedgehog signaling. Finally, we describe an efficient electroporation technique for DNA transfection into the BCC cell lines and show that they have activated Hedgehog signaling activity, albeit at a level lower than that of murine BCCs in vivo. These data indicate that the cell lines are bona fide long-term cultures of BCC cells and that DNA plasmids can be introduced into the BCC cell lines with relatively high transfection efficiency using a modified electroporation technique.