Establishment of a novel melanoma cell line SMYM-PRGP showing cytogenetic and biological characteristics of the radial growth phase of acral melanomas
Establishment of a novel melanoma cell line SMYM-PRGP showing cytogenetic and biological characteristics of the radial growth phase of acral melanomas
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DOI：
10.1111/j.1349-7006.2007.00496.x
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发表时间：
2007-07-01
期刊：
影响因子：
5.7
通讯作者：
Saida, Toshiaki
中科院分区：
文献类型：
--
作者：
Murata, Hiroshi;Ashida, Atsuko;Saida, Toshiaki
我们建立了一个新的黑色素瘤细胞系，SMYM-PRGP，这是在体内非致瘤性，从肢端黑色素瘤在放射状生长阶段在低氧环境下。对于BRAF和NRAS基因中的已知突变位点，SMYM-PRGP是野生型的，并且显示人端粒酶逆转录酶和细胞周期蛋白D1基因以及成纤维细胞生长因子-3和成纤维细胞生长因子-4基因的局灶性扩增。未观察到CDKN 2A基因突变或拷贝数丢失。p16（INK 4A）蛋白的表达水平与正常黑素细胞相同。在添加到SMYM-PRGP细胞培养物中的各种黑素细胞生长因子中，内皮素-1是最强的生长刺激剂，其效果通过添加氯化钙而显著增强。内皮素-1的生长刺激作用显示通过内皮素B受体介导。细胞周期蛋白D1在SMYM-PRGP细胞中的蛋白水平比正常黑素细胞高约10倍。虽然内皮素-1加氯化钙刺激4-6小时后增加细胞周期蛋白D1蛋白水平，磷酸化视网膜母细胞瘤蛋白的水平没有增加，这表明细胞周期蛋白D1蛋白的过度表达可能对细胞周期进程的影响不大，而是作为一个促生存因子。SMYM-PRGP是研究肢端黑色素瘤发生和进展的极好工具。
We established a novel melanoma cell line, SMYM-PRGP, which was non-tumorigenic in vivo, from an acral melanoma in radial growth phase under a low-oxygen environment. SMYM-PRGP was wild-type for known mutation sites in the BRAF and NRAS genes, and showed focal amplification of the human telomerase reverse transcriptase and cyclin D1 genes as well as the fibroblast growth factor-3 and fibroblast growth factor-4 genes. Neither mutation nor copy number loss of the CDKN2A gene was observed. The p16(INK4A) protein was expressed at a level equal to that in normal melanocytes. Among the various melanocyte growth factors added to the culture of SMYM-PRGP cells, endothelin-1 was the strongest growth stimulator, the effect of which was significantly augmented by the addition of calcium chloride. The growth stimulatory effect of endothelin-1 was shown to be mediated via the endothelin B receptor. The protein level of cyclin D1 in SMYM-PRGP cells was approximately 10 times higher than that in normal melanocytes. Although the stimulation with endothelin-1 plus calcium chloride increased cyclin D1 protein levels after 4-6 h, the level of phosphorylated retinoblastoma protein did not increase, suggesting that overexpression of cyclin D1 protein may have little effect on cell cycle progression but rather act as a pro-survival factor. SMYM-PRGP is an excellent tool for investigating the development and progression of acral melanoma.