Real-time PCR To Detect The MiR-10b,miR-126,miR-335 Express Lever In Breast Cancer And Corresponding Adjacent Normal Tissue

Objective:microRNA (miRNA) is one of RNAs, with 22 nucleotides approximately, short and noncoding, that mainly regulates the gene expression at the postranscriptional level. According to the degree of base pairing to its complementary sites within its 3'-untranslated region (3'UTR), it degrades or negatively regulates the translation of the target miRNA. Mutation, deletion or over-expression of miRNA can cause physiological anomaly or disease and correlate with various human cancers. miRNAs are involved in tumor cell proliferation, differentiation and apoptosis process, and play a role as oncogenes or tumor supressors. It is expected that microRNA might be applied in the diagnosis and treatment of Breast Cancer as an effective tool. We use real-time PCR to detect the miRNAs express lever in breast cancer and corresponding adjacent normal tissue.then predict the target genes of miRNA, Discussion the machanism in the development of breast cancer.miRNA has broad prospect in diagnosis and treatment for breast carcinoma.Methods:20 breast cancer samples and paired normal tissue samples were collected, we select the corresponding adjacent normal tissue samples which were not in the same quadrant,and were apart from the cancer at least 5cm.The total RNA was extracted from the 40 samples, reverse Transcription and real-time PCR..The results were carried on Statistical analysis.At last we use bioloinformatics methods to predict the target genes of the miRNA in the internet.Results: 1. The expression lever of miR-10b was significantly reduced in breast cancer tissue than corresponding adjacent normal tissue,the difference was Statistically significant, P= 0.0036;2. The expression lever of miR-126 was significantly reduced in breast cancer tissue than corresponding adjacent normal tissue,the difference was Statistically significant, P< 0.0001;3. The expression lever of miR-335 was significantly reduced in breast cancer tissue than corresponding adjacent normal tissue,the difference was Statistically significant, P< 0.0001;4. The relationship of the pregnancy times and the expression lever of miR-10b in normal tissue samples points that:The more women pregnancy,the higher miRNA-10b expression in the normal tissue (P=0.0069; Correlation coefficient=0.5978);5.The relationship of the pregnancy times and the expression lever of miR-335 in normal tissue samples points that:The more women pregnancy,the higher miRNA-335 expression in the normal tissue (P=O.0371; Correlation coefficient=0.59242);6. The expression lever of other miRNAs has no relationship with the age, pregnancy times,production numbers,the longest breast-feeding time,total feeding time,tumor size,TMN stage,tumor stage,lymph nodes metastasis,ER,PR,Her-2,P53,ki-67 and other factors, the difference has no Statistically significant.Conclusions:The expression lever of miR-10b,miR-126,miR-335 were significantly reduced in breast cancer tissue than corresponding adjacent normal tissue after the real-time PCR,the difference was Statistically significant,in cancer development and the process may sever as tumor suppressor gene. The more women pregnancy,the higher miR-10b and miR-335 expression in the normal tissue, pregnancy times may affect the expression lever of miR-10b and miR-335.and the expression lever of other miRNA-126 has no relationship with the clinical factors.the difference has no Statistically significant.