乙酸镍诱导16HBE细胞恶变过程中的K-ras和P15基因的改变及基因组不稳定性分析

GENOMIC INSTABILITY AND THE CHANGES OF K-ras AND 15 GENE IN TRANSFORMED 16HBE CELLS NICKEL SULFATE.

目的 :检测乙酸镍对 K- ras基因和 P15基因的改变及基因组不稳定性的影响 ,从而进一步探讨镍化合物致癌的分子机制。方法 :采用聚合酶链反应 -单链构象多态性分析方法探查乙酸镍在诱导 16 HBE细胞恶变过程中的 K- ras基因Exon1和 P15基因 Exon2存在状况。采用随机扩增多态性技术对乙酸镍在诱导 16 HBE细胞恶变过程中的基因组不稳定性进行分析。结果 :K- ras基因 Exon1和 P15基因 Exon2未发生改变。本实验所选用的 7条随机引物均能扩增出清晰、明显的条带 ,条带数在 1- 6条之间。 7条引物中 P4、P7二条引物扩增的片段在实验组和对照组之间无差异。其余 5条引物均有差异 ,对于同一随机引物他们都具有特异的带型。结论 :P15基因第 2外显子和 K- ras基因第 1外显子 (包括第 12、 13密码子 )可能不是乙酸镍作用的靶部位。在乙酸镍诱发细胞恶性转化过程中 ,基因组变得逐渐不稳定。

Objective:To detect the changes of K-ras gene and p15 gene and analyse the genomic instability in the malignant process of 16HBE cells induced by nickel acetate.Methods:PCR-SSCP was used to examine the presence of alterations of K-ras gene exon 1 and p15 gene exon 2 in the malignant process of 16HBE cells induced by nickel acetate.The genomic instability was analysed by random amplified polymorohic DNA (RAPD).Results:Compared with the negative control cells,alterations of p15 gene and K-ras gene were not observed and genomic instability was showed in the malignant process of 16HBE cells induced by nickel acetate.Conclusions:Nickel acetate maybe is not inducealteration of p15 gene and K-ras gene,indicating these four nickel compounds had not effect on the P15 gene Exon2 and K-ras gene Exon1.But they could induce genomic instability,furthermore,having special PCR bands,indicating the genome became more instably in the malignant process.