Impact Of Mir-203 On Sw620 Cells Proliferation,Migration And Invasion

Object: We tested mir-203,effects on cell activities in vitro using construction of mir-203 expression as cell proliferation?cell migration and invasion.we administered mir-203 precursor to mouse tumor xenograft model and further demonstrated colorectal cancer tumor growth in vivo.We predicted the target genes of mir-203 on the basis of the prediction of biological information through real-time PCR.the solution of questions would indicate the function of mir-203 in colorectal cancer growth and metastasis.Method:Lentiviral expression vectors LC/has-mir-203a?LV/NC were designed and constructed ? Sw620 cells were transduced with vector supernatant and subsequently through the puromycin?These two strains of cells and the lack of the transfected cell line sw620 respectively were assigned as the express group(sw620/LV-hsa-mir-203a),the negative control group(w620/LV-NC)and the blank control group.verification of continuous transfection was obtain Using micro RNA q PCR detection in 3 groups of cells.These abilities of cell proliferation,migration and invasion were asessed using cck8 method,transwell migration and invasion assay.The effect of tumorigenicity in nude mice subcutaneous was observed using the whole visualization animal model.P110 ?,AKT1 and ERL2 m RNAs were detected in three-group sw620 cell lines by q RT-PCR.Result : the results of real time q PCR shows that the sw620 cells transfected with LV-has-mir-203 expressed significantly higher than the other two control group.relative expression of Mir-203 in sw620/LVhas-mir-203 cells was 29.06±0.76,which shows statistically differences(F=4098.452,P<0.001).In contrast with the blank group,relative expression of the negative control group was 1.05 ± 0.09,and no statistical difference was builded(p>0.05).The of CCK8 cell proliferation experiment? results makes clear that the sw620 cell transfected with LV-has-mir-203 expressed significantly lower than the other control groups(F=145.243,P<0.001).there was not any statistical difference between them(p>0.05).the cck8 assay manifested that in vitro ectopic expression of Mir-203 may inhibit the proliferation of Colon tumor cells.The results of transwell chamber migration assay shows that the number-38.83±4.44 of the sw620/LV-has-mir-203 cells going through the membrane less than the blank control group and negative control group which seperate are 131.83±2.63 and 133.50±3.08(p<0.001).there was no statistical difference in the rest of these groups(p>0.05).in vitro the transwell chamber migration assay indicated that higher expression of Mir-203 can hold-up the migration of Colorectal tumor cells.The results of transwell chamber invasion assay shows that the number-10.16±1.72 of the sw620/LV-has-mir-203 cells going through the membrane less than the blank control group and negative control group(p<0.05)which seperate are 46.16±4.70 and 51.83±2.04.there was not any huge difference in the rest of these groups(p>0.05).the transwell chamber invasion assay indicated that abundant expression of Mir-203 can impress the invasion of Colorectal carcinoma cells in vitro.Cells of these groups were respectively injected Subcutaneously into the blank of nude mice.the size of nude mice,xenograft tumor was continous marked for 5 weeks from the third week.the tumour growth rate of the negative control group and blank control group were faster than that of experimental group and the difference was statistically(p<0.05).The gene' expression of p110 a,ERK2,AKT1 in sw620/LV-hsa-Mir-203 cell were less than the rest of other groups,which may be the designed genes of Mir-203.Conclusion: Mir-203 inhibited colorectal tumor sw620 cells invasion,migration and proliferation.down-regulated m RNA-expression of were found in the expressed group.mir-203 may participate in Colorectal carcinoma cell proliferation and migration through P110-alpha,AKT1 and ERK2.