Research Of MEG3Gene Expression And Methylation In Breast Cancer Tissue

Objective Long chain noncoding RNA (long, non-coding RNA lncDNA) refers to the length of more than200nucleotides, it is a noncoding RNA regulation of gene expression. Research shows that in recent years, Long chain noncoding RNA affect the growth of tumor cells by epigenetic regulation.. Through the analysis of long-chain MEG3noncoding RNA genes in breast cancer and the expression level of Paracancerous Normal Tissue, Application of methylation specific PCR (methylation specific PCR, MSP) technology. Analysis MEG3gene sequences of DNA methylation,dicuss the relationship between methylation of MEG3and its expressiom.Methods Collection of breast cancer and adjacent normal tissues of20normal tissues adjacent to cancer, select non same quadrant, from the tumor tissue mass is at least more than5cm tissue of breast cancer tissue. Extract total RNA,,determination of purity and integrity,the MEG3gene expression was detected by real-time fluorescence quantitativePCR.Take the10on breast cancer and5paracancerous 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.Result1. The expression level of Meg3in tissues of breast cancer and adjacent normal tissue. By real-time PCR detection, the expression of MEG3gene in breast carcinoma tissue was significantly lower than that in normal tissue expression, there were statistically significant differences, P=0.00.2.Detect DNA methylation of MEG3gene in breast cancer tissue and adjacent tissue.There are9cases of breast cancer tissues appearing methylation of MEG3DNA,non-methylation lcase;while methylation in tissue adjacentto breast carcinoma with2cases,and3cases have no methylation,P=0.07.3.Effects of MEG3gene methylation in breast cancer and adjacent tissue to the expression of MEG3.IN all samples,there are11cases appear methylation,4cases without methylation,P=0.07.]Conclusion By real-time PCR,we found the expression level of MEG3gene significantly downregulated in breast carcinoma tissue than adjacent normal breast tissue, the difference was statistically significant (P<0.05).The result similar to the report both at home and abroad.By means of MSP for analysis MEG3DNA methylation status of breast cancer and normal adjacent tissue,the methylation status has no significant difference,there is no statistically significant(P>0.05).Different methylation status in the transcription and expression of MEG3in breast cancer and normal breast adjacent tissue,has no significant difference, there is no statistically significant(P>0.05).The result may declare that in the occurrence and development of breast cancer there is’t regulation by DNA of MEG3methylation.