A Study Of Hsa-miR-363、Hsa-miR-4-23-5p Regulating The Expression Of VEGF In HaCaT Cells

Objective:To explore the regulative effect of expression of VEGF gene in the hsa-miR-363and hsa-miR-423-5p,and to discuss the future application of hsa-miR-363and hsa-miR-423-5p in the gene therapy for psoriasis.Methods:HaCaT cells were cultured in vitro-Planking training after the completion of the cells.There were seven groups:hsa-miR-363mimic group、 hsa-miR-363inhibitor group-. hsa-miR-423-5p mimic group、hsa-miR-423-5p inhibitor group、mimic NC group、inhibitor NC group and blank control group.First, adding transfection reagent for transfection. Second, with determined by MTT method to detect HaCaT cell proliferation activity and cell transfection. The VEGF mRNA and VEGF protein were detected by real time reverse transcriptase polymerase chain reaction(qRT-PCR) and ELISA respectively. Levels of VEGF in hsa-miR-363and hsa-miR-423-5p were measured using quantitative sandwich enzyme-linked monoclonal immunosorbent assay.Results:hsa-miR-363mimic group、hsa-miR-363inhibitor group hsa-miR-423-5P mimic group、hsa-miR-423-5P inhibitor group、mimic NC group、 inhibitor NC group and blank control group determined by MTT experiment result display cell proliferation activity was statistically significant (P<0.05).We can see the cell grow well In the microscopic and on behalf of transfection effect also is very good. Cellular proliferation inhibition rate reached the highest at72h. It illustrate the transfection efficiency was highest. The highest transfection efficiency reflect obvious difference between the groups. RT-PCR showed that hsa-miR-363mimic and hsa-miR-363inhibitor、hsa-miR-423-5P mimic and hsa-miR-423-5P inhibitor had no influence on the expression of VEGF gene mRNA. ELISA result display each of the hsa-miR-363mimic group、hsa-miR-363inhibitor group、mimic NC group、 inhibitor NC group and blank control group were statistical significance at VEGF protein expression levels. But each of the hsa-miR-423-5P mimic group、 hsa-miR-423-5P inhibitor group、mimic NC group、inhibitor NC group and blank control group were no statistical significance.Conclusion:Hsa-miR-363makes the mRNA of VEGF in the HaCaT cell overexpressed.Hsa-miR-423-5P has no effect on the expression of VEGF. Psoriasis is a disease of genetic abnormalities, the hsa-miR-363can regulate a large number of mRNA negativly on the level of transcription, the regulated mRNA are interacted between each other, and format a network of co-regulation of the structure by a variety of ways to participate in the occurrence of psoriasis.