Oncogene mutation analysis that uses malignant melanoma model mouse and mRNA analysis by ISH method.

使用恶性黑色素瘤模型小鼠进行癌基因突变分析，并通过 ISH 方法进行 mRNA 分析。

• 批准号: 20790822
• 负责人: YAMAZAKI Fumikazu
• 金额: $ 2.66万
• 依托单位: Kansai Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2008
• 资助国家: 日本
• 起止时间: 2008 至 2009
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-20790822/
• 关键词: 遺伝子; 悪性黒色腫; サイトカイン; ISH法; 癌

We analyzed the mutation and methylation of p16 tumor suppressor gene, using UVB induced melanoma in XPA-deficient (-/-), stem cell factor-transgenic (SCF-Tg) mice. Method : Genomic DNA was extracted. The full lengths of exon 1 and exon 2 were amplified by PCR, followed by the direct sequencing and methylation specific PCR. Results and discussion: The complete deletion of p16 gene was observed in 5 of 10 (50%) NM samples, and no point mutation was observed in other NM samples. No mutation was observed in any sample of LMM, SCC, or chronically UVB-irradiated skin. DNA methylation was detected in 3 of 10 (33%) in NM samples and 1 of 14 (7%) in LMM samples. These results suggest that the deletion and methylation of p16 gene would lead to the carcinogenesis of NM type in our model mouse, and the factor(s) other than p16 gene mutation would be involved in the carcinogensis of LMM type.

我们分析了p16抑癌基因的突变和甲基化，利用UVB诱导XPA缺陷（-/-），干细胞因子转基因（SCF-Tg）小鼠的黑色素瘤。方法：提取基因组DNA。PCR扩增外显子1和外显子2的全长，然后进行直接测序和甲基化特异性PCR。结果和讨论：10例NM中5例（50%）p16基因完全缺失，其余NM未见点突变。在任何LMM、SCC或慢性UVB照射皮肤样本中均未观察到突变。在NM样品中的10个中的3个（33%）和LMM样品中的14个中的1个（7%）中检测到DNA甲基化。提示NM型小鼠的癌变可能与p16基因的缺失和甲基化有关，LMM型小鼠的癌变可能与p16基因突变以外的因素有关。