人胰腺腺癌细胞系中p16基因、cyclin D1基因和pR6蛋白的改变

目的：研究胰腺腺癌细胞系ｐＲｂ－ｐ１６细胞调节途径相关因子的异常情况。方法：用ＰＣＲ及单链构象多态性（ＳＳＣＰ）的方法检测５株人胰腺腺癌细胞系ｐ１６基因的缺失和突变;用Ｓｏｕｔｈｅｒｎ印迹分析ｃｙｃｌｉｎ Ｄ１基因的扩增;用Ｗｅｓｔｅｒｎ印迹分析检测ｐＲｂ基因的表达。结果：２株胰腺腺癌细胞系存在ｐ１６基因第１外显子纯合性缺失,未发现第１、２外显子的基因突变;１株细胞系有ｃｙｃｌｉｎ Ｄ１基因的扩增;ｐＲｂ基因在５株细胞系中均呈高磷酸化的表达。结论：胰腺腺癌细胞系中存在ｐＲｂ－ｐ１６细胞调节途径的异常,主要表现为ｐ１６和ｃｙｃｌｉｎ Ｄ１基因的改变。

Changes of p16 gene,cyclin D1 gene,and pRb protein in human pancreatic adenocarcinoma cell lines

OBJECTIVE: To investigate the abnormalities of the relative factors of pRb-p16 cell regulatory pathway in pancreatic adenocarcinoma. METHODS: Five strains of human pancreatic adenocarcinoma cell lines were analysed by PCR and single-strand conformation polymorphism (SSCP) for homozygous deletion and mutation of the p16 gene, respectively. The gene amplification of cyclin D1 was detected by Southern blot analysis and the protein expression of pRb gene was examined by Western blot analysis. RESULTS: Homozygous deletion of exon 1 were found in two cell lines and no mutation was found within exon 1 and exon 2 in five cell lines. Cyclin D1 gene amplification was observed in one cell line and all of the five cell lines showed hyper-phosphorylated pRb protein. CONCLUSION: These findings have demonstrated that there are abnormalities of pRb-p16 regulatory pathway in pancreatic adenocarcinoma cell lines, revealing the alteration of the p16 and cyclin D1 genes.