| Objective :To investigate the effects of 5-fluorouracil( 5-FU) and Trichostatin A( TSA) on biological behavior of colorectal cancer Lovo cell lines, and probe the influence TSA on chemotherapy resistance of 5-FU as well as the possible mechanisms in Lovo cells.Methods :Colorectal cancer Lovo cell line was chosen to experiment, and the experiments included five groups: control group, 5-FU group, TSA group, TSA preconditioning group and combination group( 5-FU + TSA).(1)Using methyl thiazolyl tetrazoliu( MTT) colorimetric assay and soft agar colony formation to observe cell proliferation of each group.(2)Invasion and migration capacity used transwell tumour cell invasion assay and scratch healing experiment after drugs treatment 24 h.(3)Flow cytometer was used to observe apoptosis rate and cell cycle of each group cells after drugs intervention 24 h.(4)The expression of thymidylate synthase( TS) of each group cells was detected by Western-blot after drugs treatment 24 h.Results :(1)MTT cell proliferation assay: in contrast with control group, the 5-FU group, TSA preconditioning group and combination group had a growth inhibition at 24 h, 48 h and 72h( p < 0.05), and TSA group presented a growth inhibition at 48 h and 72 h( p < 0.05). In comparison with 5-FU group, the growth inhibition of TSA preconditioning group and combination group were distinctive at 48 h and 72 h( p < 0.05), but the inhibition between TSA preconditioning group and combination group were non-significant( p > 0.05).(2)Soft agar colony formation assay: in comparison with control group, each experimental group colony formation number was markedly reduced( p < 0.05).Compared with 5-FU group, the number of colony formation in TSA preconditioning group and combination group significantly decreased( p < 0.05).(3)Invasion and migration ability assay: compared with control group, penetrating the matrigel cell number and scratch healing rate in the experimental groups significantly reduced( p < 0.05). Compared with 5-FU group, penetrating the matrigel cell number and scratch healing rate in TSA preconditioning group and combination group notably decreased( p < 0.05), but the cell number penetrating the matrigel and scratch healing rate between TSA preconditioning group and combination group were non-significant( p >0.05).(4)Apoptosis assay: in contrast to control group, the apoptosis rate of the experimental groups significantly increased( p < 0.05), in compare with 5-FU group, the apoptosis rate of TSA preconditioning group and combination group notably incresaed( p < 0.05), but the apoptosis rate between TSA preconditioning group and combination group were non-distinctive( p > 0.05).(5)Cell cycle assay: compared with control group, the drug-treatment groups in G0/G1, S and G2/M periods had no significance( p > 0.05).(6)TS expression assay: the differs of TS expression in control group and 5-FU group were not significant( p > 0.05), compared with control group, TS expressions of TSA group, TSA preconditioning group and combination group were notably decreased( p <0.05). Compared with 5-FU group, TS expressions of TSA group, TSA preconditioning group and combination group were markedly decreased( p < 0.05), but TS expressions of the three groups were non-significant( p > 0.05).Conclusion :(1)5-FU and TSA can inhibite proliferation, invasion and migration of Lovo cells, and increase their apoptosis.(2)TSA can increase the chemotherapy sensibility of 5-FU in Lovo cells to some extent, the possible mechanism is reducing the TS expression. |
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