重铬酸钾致N-ras基因DNA损伤作用研究

Detecting the DNA damage of N-ras gene induced by potassium dichromate

目的 研究重铬酸钾对N -ras基因的DNA损伤作用,探讨其致癌作用分子机制。方法 制备N -ras基因外显子1单链探针;重铬酸钾染毒细胞提取基因组DNA ,再经RDPCR扩增后与探针杂交显色。结果 重铬酸钾染毒剂量10 0 μmol L可检测到DNA损伤片段杂交条带,其损伤片段长于完整的N ras基因外显子1;更高剂量(10 0 0 μmol L)未能检测出DNA损伤作用。结论 重铬酸钾可能造成N- ras基因外显子1上游序列DNA损伤,且该损伤作用可能正是其致癌作用分子机制的关键所在。

Objective Applying RDPCR to detect the DNA damage of N-ras gene induced by Potassium dichromate Methods Preparing single-stranded probes of exon 1 of N-ras gene in human. The genomic DNA was treated with Potassium dichromate,then amplified by RDPCR and detected by Southern hybridization with the probe. Results The clear hybridized bands can be seen in the position which is induced by Potassium dichromate on the dose of 100μmol/L, but can't be detected on the dose over 1000μmol/L Conclusion It indicates that Potassium dichromate can cause the DNA damage of N-ras gene, which should be the key point of its carcinogenesis mechanisms.