Effect Of MicroRNA-145 On The Proliferation And Apoptosis Of Cutaneous Basal Cell Carcinoma Cells

Objective: To investigate the effects of miRNA-145 on the proliferation、apoptosis and cell cycle of cutaneous basal cell carcinoma A431 cells.Methods: Taking the cutaneous basal cell carcinoma A431 cell line as the research object,miR-145 mimics and negative control sequence(NC mimics)were transfected into the cutaneous basal cell carcinoma A431 cells with liposome transfection method.The experiment was divided into 3 groups: miR-145 group(transfection miR-145mimics),NC group(transfection negative control sequence)and Mock group(no transfection of any sequence),observing the transfection efficiency under a fluorescence microscope after transfection.The expression level of miR-145 in the 3groups of cells was detected by Real-time PCR;The proliferation of the 3 groups of cells was detected by CCK-8 experiment;The cell cycle and apoptosis of the 3 groups of cells was detected by flow cytometry;Western blotting was used to detect the expression of caspase-3 protein in 3 groups of cells.result:1.Real-time PCR results showed that the expression of miR-145 in the miR-145 group was 3.76±0.28,which was significantly higher than the NC group(1.00±0.10)and Mock group(1.00±0.10),and the difference was statistically significant(P<0.01).The difference between NC group and Mock group was not statistically significant(P>0.05).2.The results of the CCK-8 experiment showed that the absorbance value of the cells in the miR-145 group was 0.66±0.02,which was significantly lower than the NC group(0.91±0.05)and Mock group(0.93±0.03),and the difference was statistically significant(P <0.01).The difference between NC group and Mock group was not statistically significant(P>0.05).3.The apoptosis results showed that the total apoptosis rate of the miR-145 group was 26.26%±1.25%,which was significantly higher than that of the NC group(5.82%±0.21%)and Mock group(5.54%±0.38%),and the difference was statistically significant(P<0.01).The difference between the NC group and the blank control group was not statistically significant(P>0.05).4.The cell cycle results showed that the proportion of G0/G1 phase in the miR-145 group was 87.52%±0.72%,which was significantly higher than the NC group(61.65%±0.81%)and Mock group(62.09%±0.47%),and the difference was statistically significant(P<0.01);The proportion of S phase in the miR-145 group was 3.57%±0.70%,which was significantly lower than the NC group(31.66%±0.31%)and Mock group(31.27%±1.15%),and the difference was statistically significant(P<0.01).There was no significant difference in the proportion of cells in G0/G1 phase and S phase between the NC group and Mock groups(P>0.05).5.The results of the Western-blotting experiment showed that the expression of Cleaved-caspase-3 protein in the miR-145 group was 0.37±0.02,which was significantly higher than the NC group(0.12±0.04)and Mock group(0.12±0.03),and the difference was statistically significant.The difference between the NC group and Mock group was not statistically significant(P<0.01);There was no significant difference in the expression of Procaspase-3 protein among the three groups(P>0.05).Conclusion: 1.Overexpression of miR-145 can inhibit the proliferation of cutaneous basal cell carcinoma A431 cells、induce cell apoptosis、changes the cell cycle distribution and promote the cleavage of caspase-3 protein.2.miR-145 may play an important role in the proliferation、apoptosis induction and cell cycle regulation of cutaneous basal cell carcinoma cells.3.miR-145 may act as a tumor suppressor gene in the cutaneous basal cell carcinoma and is a potential therapeutic target for the cutaneous basal cell carcinoma.