摘要
目的 检测结晶型硫化镍 (NiS)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 采用限制性片段长度多态性分析和聚合酶链反应 -单链构象多态性分析方法探查结晶型NiS在诱导 16人气管上皮细胞 (HBE)恶变过程中的K -ras基因Exonl第 12密码子及第 12密码子以外的密码子改变情况。采用PCR -SSCP分析方法探查结晶型NiS在诱导 16HBE细胞恶变过程中的P15基因Exon2存在状况。采用随机扩增多态性技术来对结晶型NiS在诱导 16HBE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。其中P4、P5、P7两条引物扩增的片段在实验组和对照组之间无差异 ,其余 4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 )可能不是结晶型NiS作用的靶部位。在结晶型NiS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 alterations of k-ras gene and p15 gene and analyse the genomic instability in the malignant process of 16HBE cells induced by crystalline nickel sulfide(NiS).Methods PCR-RFLP was used to detect the mutation of 12 code of k-ras gene and 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 crystalline nickel sulfide.Analysing the genomic instability 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 crystalline nickel sulfide.Conclusion Crystalline nickel sulfide could not induce alteration of p15 gene and k-ras gene,which indicated these four nickel compounds had not effect on the P15 gene exon2 and K-ras gene exonl,but they could induce genomic instability,furthermore,having special PCR bands,indicating genome became more instability in the malignant process.
出处
《中国公共卫生》
CAS
CSCD
北大核心
2004年第7期777-779,共3页
Chinese Journal of Public Health
基金
国家自然科学基金项目 (391 70 651 )
