Research Of HOTAIR Gene Expression And Methylation In Breast Cancer Tissue

Objective:By analyzing the Long non-coding RNA genes HOTAIE expression levels in breast cancer and adjacent tissues. Methylation-specific PCR (methylation-specificPCR, MSP) technology, analysis of the methylation status of breast cancer and adjacent normal tissues HOTAIE genomic DNA. Explore the relationship between DNA methylation and its HOTAIE gene expression.Methods:Collection of breast cancer and adjacent normal tissues of 30 normal tissues adjacent to cancer, select non same quadrant, from the tumor tissue mass is at least more than 5cm tissue of breast cancer tissue. Extract total RNA, determination of purity and integrity,the HOTAIR gene expression was detected by real-time fluorescence quantitative PCR.Take the 30 on breast cancer and paracancerous tissue respectively for genomic DNA extraction.Then treat with bisulfite,design of methylated and non methylated primerscorresponding series parallel PCR amplification. Through Agarose gel electrophoresis the analysis, to determine the methylation status of DNA, The data were statistically analyzed.Results:1.by the paired T test was used for statistical analysis, the real time fluorescent quantitative detection of breast cancer and matched normal tissues we found:the expression level of HOTAIR was 11.16±3.97 in breast cancer, the expression of HOTAIR in tumor adjacent normal tissues was 5.02±2.58, P=0, P<0.05, the difference was statistically significant.2. After MSP test, we found that HOTAIR in breast cancer tissue and 27 cases of nonmethylation, methylation in 3 cases; and the adjacent tissues in 17 patients with out methylation, the methylation of in 13 cases, P=0.004 P<0.05,the difference was statistically significant.Conclusions:1. Detection of HOTAIR expression in 30 cases of breast tissue activity; in breast cancer tissues, the expression of HOTAIR was significantly higher than that in normal tissues,the difference was statistically significant (P=0.000). These results suggest that high expression of HOTAIR may be associated with the occurrence of breast cancer2. In the HOTAIR visible in 30 cases of breast cancer tissues and adjacent normal tissuesgene promoter CpG island methylation level is higher than that of non cancerous tissue,and the statistical analysis confirmed that the difference between the two was statistically significant (P=0.004). The above results suggest:DNA methylation may be one of the reasons for the high expression of HOTAIR gene in breast cancer tissues.3. the experimental results found, non methylation status of HOTAIR gene promoter CpG island and the gene is highly expressed in breast cancer, may play an important role in carcinogenesis of breast cancer. The experimental results provide a theoretical basis for the specific indicator of new breast cancer detection for HOTAIR gene methylationlevel to become, and may become the specific biological potential target for treatment of breast cancer. But due to the HOTAIR promoter region with multiple methylated regions, will also need to study quantitative and qualitative analysis of multiple regions, in order to determine the specific relationship between HOTAIR methylation and breast cancer...