ABSTRACT
Objective: To verify whether EFNB2 gene is targeted by miRNA-497 using molecular biology methods. Methods: Bioinformatic method predicted that EFNB2 gene is targeted by miRNA-497. PCR methods amplified the fragment in EFNB2 gene 3'-UTR including the putative miRNA-497 binding site. Then the sequences of wide type (WT) and mutant (MUT) were cloned into the pmirGLO luciferase vector, respectively. The DNA sequences of the amplified fragments were identified by restriction enzyme digestion and sequencing, and were consistent with the reference sequence from UCSC. This constructed vector was marked as pmirGLO-EFNB2 vector. Finally, the pmirGLO vector, the pmirGLO-EFNB2 vector, miRNA-497 mimics and negative control (NC) were divided into five groups and transfected into HEK393T cells, and the luciferase activity was tested after 24 h and 48 h by dual luciferase reporter gene assay. Results: The results of DNA sequencing demonstrated that the PCR fragment was successfully cloned into pmirGLO vector. The transfection results showed that the recombinant plasmid of WT and MUT was successfully transfected into HEK293T. The results of dual luciferase activity assay demonstrated that miRNA-497 significantly decreased the reporter gene activity compared with the NC. Conclusion: At the cellular level, the schizophrenia susceptibility gene EFNB2 was verified to be targeted by miRNA-497, which provides a new idea and new clue for subsequent studies on miRNA-497 function in the molecular mechanism of schizophrenia.
ABSTRACT
Objective To investigate the value of IL-27,IL-29 and miRNA-497 in HER-2 positive radioactive particles combined targeted therapy. Methods Sixty-one elderly inpatients with HER-2 positive gastric cancer ascites in the Gaoming District People's Hospital, Wuchuan Municipal People's Hospital and Zhejiang Provincial Tumor Hospital were divided into the two groups. The treatment group(n= 31) was treated by the radioactive particles+ targeted drug Trastuzumab(initial dose 8 mg/kg, maintenance dose 66 mg/kg, once per 3 weeks) for 6 weeks; the control group(30 cases) was treated by targeted drug Trastuzumab (initial dose 8 mg/kg,maintenance dose 6 mg/kg, once per 3 weeks) for 6 weeks. The levels of ascites IL-27,IL-29 and miRNA-497 in the radioactive partcles combined targeted therapy group and control group were detected. Results The levels of ascites IL-27, IL-29 and miRNA-497 after treatment in the treatment group were significantly decreased compared with those before treatment and in the control group,the effective remission rate in the treatment group was 74. 19%, which was higher than 36.67% in the control group, the average median survival time in the treatment group was 155 d, which washigher than 72 d in the control group, the difference was statistically significant. Conclusion IL-27 ,IL-29 and miRNA-497 has the significance of curative effect evaluation and prognosis judgment in the radioactive particles combined targeted therapy for the patients with HER 2 positive gastric cance.