Detection Of PANDAR Gene DNA Methylation In Breast Cancer By COBRA And QM-PCR

Objective:To quantitatively detect and analysis the DNA methylation of PANDAR gene in breast cancer tissues and the adjacent normal tissues by combined bisulfite restriction analysis (COBRA) and QM-PCR. In order to explore the clinical significance of the DNA methylation of PANDAR gene in breast cancer.Methods:The breast cancer tissues and the adjacent normal tissues (no less than 5 cm away from the lumps) collected from different quadrants (n=30) were extracted DNA respectively, and quantitatively detect and analysis the DNA methylation of PANDAR gene by COBRA and QM-PCR. The statistical analysis was performed with SPSS 17.0.Results:1. The results of COBRA showed in the 30 cases digested by Bsp1191 enzyme, the DNA methylation degree of PANDAR gene was above 50%in 28 cases, and below 50%in two cases; in the cases digested by Hinfl enzyme, the DNA methylation degree was above 50%in 30 cases, and none was below 50%. There was no significant difference (p= 1> 0.05).2. The results of fluorescence quantitative QM-PCR showed the DNA methylation rate of PANDAR gene was 74%in the 30 breast cancer tissues and the adjacent normal tissues, and was 68%in the adjacent normal tissues. There was a significant difference (p= 0.0069> 0.05).Conclusion:1. The promoter CpG island methylation degree of PANDAR gene in the 30 breast cancer tissues and adjacent normal tissues was higher than that in the adjacent tissues. These results suggested that there was a correlation between DNA methylation and the higher expression of PANDAR gene in breast cancer tissues.2. The DNA methylation degree of PANDAR gene in breast cancer tissues was higher than that in adjacent normal tissues. There was a correlation between the promoter CpG island methylation status of PANDAR gene and the higher expression of PANDAR gene in breast cancer tissues. It was suggested that the DNA methylation of PANDAR gene is likely to be a valuable molecular markers and a new specific indicator for detecting breast cancer., and might play an important role in the initiation and progression of breast cancer, implicating as a potentially specific therapeutic target for breast cancer.