Invasiveness Of Human Hepatoma Cell Line SMMC-7721 After Changing Cell Cycle Phase Distribution

Objective:1.Observe the cell cycle phase distribution of human hepatoma cell line SMMC-7721 after CDKs was inhibted.2.Observe the invasiveness of human hepatoma cell line SMMC-7721 after the cell cycle phase distribution was changed。3.Elucidate the correlationship of the invasiveness with cell cycle phase distribution.Method:1.Cells are devided into 2 groups(group A: treatmented with Roscovitine 0umol/L for 24h, group B: treatmented with Roscovitine 32μmol/L for 24h),The cell cycle phase distribution of 2 groups is detected by flow cytometry analysis.2.The motor ability and invasiveness of 2 groups is studied by wound healing assay and transwell chamber assay.3.The mRNA expression of uPA and MMP-9 of 2 groups is studied by RT-PCR.Results:1.The proportion of group B in G0/G1 phase is increased compear with group A (72.19±0.47%vs59.22±0.54%, P<0.05), in S phase is decreased (18.95±0.24% vs 26.34±0.20%,P<0.05),in G2/M phase is decreased(8.86±0.76% vs 14.44±0.62 %,P<0.05).2. The motor ability in group B is decreased(3.52±2.35 vs14.02±2.93,P<0.05);the number of cell penetrated membrance is decreased (71.40±5.59vs149.60±16.36,P<0.05) ,after most human hepatoma cell line SMMC-7721 cells were blocked in G0/G1 phase, the invasiveness is decreased.3.RT-PCR analysis reveal that expression level of uPA mRNA is decreased ,but the expression level of MMP-9 mRNA is no significantly change.Conclusion: 1.Most cells of human hepatoma cell line SMMC-7721 are blocked in G0/G1 phase after CDKs activity was intervented by Roscovitine.2. The invasiveness could be inhibited after human hepatoma cell line SMMC-7721 cells were blocked in G0/G1 phase.3. The decline of invasiveness may be relate with the change of motor ability and down-regulation of uPA expression level.