Effects Of Cofilin-1on Proliferation And Migration Activity In Human Skin SCC A431Cells

Objective:In this study, we investigate the effect of silencing cofilin-1gene expression andbaicalin on cell proliferation and cell migration in human skin SCC A431cell line.Methods:1. Cultured cells: in high glucose-DMEM with10%fetal bovine serum FBS culturemedium of human skin squamous cell carcinomaA431cells at37℃,5%CO2incubator2. Drug deployment: prepare baicalin with1ML dimethyl sulfoxide (DMSO) as asolvent, the deployment of a concentration of40mg/ml, kept in a brown container into therefrigerator-20℃to spare. In this experiment, the final concentration of baicalin was50μg/ml.3. siRNA interference: For cofilin-1mRNA sequences were designed andsynthesized specific siRNA, was transfected into A431cells, and in a certain time to join50μg/mL baicalin, using real-time quantitative PCR (Real-time PCR) to detect cofilin-1gene silencing.4. Cell proliferation assay: The A431cell proliferation was detected by CCK8.5. Cell migration assay: scratch wound healing assay.Results:1. According to the RT PCR results, the expression of cofilin-1mRNA were lowerafter siRNA interference than the control group, the difference was statisticallysignificance (P <0.05), showed significant inhibition of cofilin-1expression..Therefore, inthe following experiment, we choose the best silencing effect of siRNA fragments.2. And comparing the control group,the cell proliferation rate were significantly decreased,for24h,48h,72h the A431cell were transfected with cofilin-1siRNA; when thebaicalin added, A431cell proliferation have a significantly reduced, whereas the baicalintreatment and cofilin-1siRNA together,the celluar proliferation rates were the mostsignificantly reduced,with time extending,the cell activity has not enhanced;this indicatingthat the proliferation of A431cells after gene silencing cofilin-1was significantly reduced,baicalin treatment can also dampen the proliferation rate, however, cofilin-1gene silencingsignificantly increased the effect of baicalin;3.The result of scratch wound healing assay showed:Single baicalin treatment andcofilin-1silencing could dreased the A431cell growth and migration. And cofilin-1silencing significantly enhancing the efficacy of baicalin.Conclusion:This study showed that cofilin-1silencing results in significant inhibition of tumorgrowth and migration in the A431cell,Single baicalin treatment could significantly inhibitthe tumor cells growth and migration in the A431cell line.And cofilin-1silencingsignificantly enhancing the efficacy of baicalin. And cofilin-1meight become the potentialtarget gene to enhance the effect of anticancer drugs.