The Screening Of MiR-210Target Gene And The Research In Mechanism In Breast Cancer

Objective:The target genes of miR-210were achieved in prior research by prediction software, such as MEF2A, EGR3, SETD2, E2F3, CHAD, AIFM3, HMGB1. In order to supply new target for diagnosis and treatment in breast cancer, this research fartherly prove seven prediction gene by real-time PCR and Western blot, and preliminary analysis the mechanism of these genes in breast cancer.Methods:miRNA-210mimic, miRNA-NC, miR-210inhibitor, miR-210-INC was transfected to human breast cells MDA-MB-231by lipofectamineTM2000to reduce or inhance the activity of miR-210and the fluorescence microscope was used to detect the transfection efficiency. The relative expression of miR-210was detected by real-time PCR24h,48h and72h after transfection. Total RNA was abstracted after transmit by TROZIO, and was detected its quality by agorose, and was reversed into cDNA. Target genes were amplified via cDNA template by real time PCR.Results:1MiRNA-210mimic, miRNA-NC (20pmol), miR-210inhibitor and miR-210-INC were successfully transmited into MDA-MB-231cells, transfection efficiency. high transfection efficiency is (87.31±2.68)%. the content of miR-210inhibitor transmited after48hours is lowest, for The inhibition of miR-210inhibitor to the miR-210is best.2. Optical density (OD) and concentration, of total RNA was deteted by ultraviolet specrophotometer:A260/A280=1.8-2.0. concentration of total RNA is300-700μg/ml. The appearances of5S band, clear28S and clear18S suggested that RNA was complete after the sepharose electrophoresis of RNA extracted.3. The expression of MEF2A, EGR3, SETO2, CHAD, HMGB1, AIFM3have no statistically significant, whereas the expression of EGR3have remarkablely statistically significant.4. EGR3protein bands were enhanced, and have remarkablely statistically significant compared to control (P<0.05).Conclusion:1. MEF2, EGR3, SETD2, CHAD, HMGB1, AIFM3are not target gene, and EGR3may be its target genes.2. Above Researches Play a important role in exploring mechanism and provide a thinking for the diagnose and treatment for breast cancer.