The Effect Of MiRNA-145 On Proliferation And Apoptosis Of Cutaneous Squamous Cell Carcinoma Cells

Objective:To investigate how mi RNA-145(mi R-145)affects the proliferation,cycle,and apoptosis of cutaneous squamous cell carcinoma A431 cells.Methods:Cutaneous squamous cell carcinoma cell A431 cells were transfected with mi R-145 mimic and mi R-145 negative control plasmid via liposome(Lipo2000).The experiment was divided into three groups:mi R-145 mimic transfection group,negative plasmid transfection group and blank control group(without any sequence).The transfection efficiency of the three groups was observed under fluorescence microscope after transfection.The changes in the expression of mi R-145 in the three groups were verified by q RT-PCR,CCK-8 was used to verify the changes in the proliferation of the c SCC A431 cells,flow cytometry was used to confirm changes of the percentage of the cell cycle and apoptosis in c SCC A431 cells and western blotting was performed to identify the changes of the production of Caspase-9 and Caspase-3 in c SCC A431 cells.Results:1.there are about 70% of the cells in the mi R-145 mimic transfection group and the negative plasmid transfection group can be seen green fluorescence expression under the fluorescence microscope,while the cells in the blank control group can’t be seen green fluorescence expression.2.In the mi R-145 mimic overexpression group,the expression levels of mi R-145 was(2.95±0.10)was significantly higher than the negative plasmid transfection group(1.10±0.14)and the blank control group(1.00±0.11)(p<0.01);the plasmid transfection group and the blank control group did not show any significant(p>0.05).3.The OD value of the mi R-145 mimic transfection group,the negative plasmid transfection group and the blank control group after 24 h were respectively(1.15±0.01)、(1.19±0.01)、(1.20±0.02),the OD value of the three groups after 48 h were respectively(1.22±0.01)、(1.35±0.01)、(1.38±0.02),the OD value of the three groups after 72 h were respectively(1.43±0.01)、(1.69±0.004)、(1.71±0.01).The cell proliferation level of the mi R-145 mimic transfection group was significantly lower at24 h 、 48 h 、 and 72 h when compared to the the blank control group and negative plasmid transfection group(p<0.01).There would be no statistically significant difference between the blank control groups and plasmid transfection(p>0.05).4.The proportion of cells in G0/G1 phase of the mi R-145 mimic transfection group[(87.5±0.50)%]was significantly higher than the negative plasmid transfection group[(61.60±0.35)%] and the blank control group [(61.90±0.39)%](p<0.01),the percentage of cells in S phase of thhe mi R-145 mimic transfection group[(3.46±0.43)%] was significantly lower than the negative plasmid transfection group[(31.51±0.23)%] and the blank control group [(30.85±0.61)%](p<0.01),The deference between plasmid transfection group and the blank control group did not vary significantly(p>0.05).5.The apoptosis rate of mi R-145 mimic transfection group [(34.16±0.31)%] was higher than the negative plasmid transfection group[(13.92±0.25)%] and the blank control group [(14.13±0.20)%](p<0.01),The deference between plasmid transfection group and the blank control group did not vary significantly(p>0.05).6.The relative expression levels of Caspase-9 in the mi R-145 mimic transfection group(1.36±0.03)was significantly higher than the negative plasmid transfection group(0.54±0.03)and the blank control group(0.53±0.05)(p<0.01),the relative expression levels of Caspase-3 in the mi R-145 mimic transfection group(1.44±0.05)was significantly higher than the negative plasmid transfection group(0.63±0.02)and the blank control group(0.60±0.02)(p<0.01),while there was no difference between the negative plasmid transfection group and the blank control group(p>0.05).Conclusion:1.mi R-145 expression can inhibit the proliferation and induce apoptosis of cutaneous squamous cell carcinoma A431 cells,which may inhibit the occurrence and development of cutaneous squamous cell carcinoma.