Hsa-miR-30e、Hsa-miR-192Regulate The Expression Of Vascular Endothelial Growth Factor In Human HaCaT Keratinocytes

Objective:To explore the regulative effect of expression of VEGF gene in the hsa-miR-30e and hsa-miR-192,and to discuss the future application of hsa-miR-30e and hsa-miR-192in 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-30e mimic group、hsa-miR-30e inhibitor group、hsa-miR-192mimic group、hsa-miR-192inhibitor 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-30e and hsa-miR-192were measured using quantitative sandwich enzyme-linked monoclonal immunosorbent assay.Results:hsa-miR-30e mimic group、hsa-miR-30e inhibitor group、hsa-miR-192mimic group、hsa-miR-192inhibitor 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.There was no significant difference in the hsa-miR-192mimic group and hsa-miR-192inhibitor group at24h. We guess may be little difference between the two group. Other cell proliferation inhibition rate between groups was statistically significant.48h, hsa-miR-30e mimic and hsa-miR-30e inhibitor in the two groups compared with the blank control group in cell proliferation inhibition rate was statistically significant. Hsa-miR-192mimic, hsa- miR-192inhibitor and blank each cell proliferation inhibition rate between the control group (P<0.05). It would have a little fluctuation between groups with the transfection of the effect was more apparent.72h, only hsa-miR-30e mimic and hsa-miR-30e inhibitor, hsa-miR-192mimic and hsa-miR-192inhibitor compare cell proliferation inhibition rate has no statistical significance.The highest transfection efficiency reflect obvious difference between the groups.qRT-PCR result display the value of hsa-miR-30e mimic was (6.04±0.07). That raised the hsa-miR-30e abundance of VEGF gene mRNA expression is higher than normal VEGF gene mRNA expression (6.04±0.07) times. Hsa-miR-192mimic and hsa-miR-192inhibitor of VEGF gene mRNA expression was not detected. ELISA result display each of the hsa-miR-30e mimic group、hsa-miR-30e inhibitor 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-192mimic group、hsa-miR-192inhibitor group、mimic NC group、inhibitor NC group and blank control group were no statistical significance.Conclusion:Psoriasis is a disease of genetic abnormalities, the hsa-miR-30e can 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.