Study On Treatment Of The ERK_1/2 Inhibitor Combined With 5-FU For Malignant Melanoma

Objective:This study was to observe the effects of combination of the ERK1/2 inhibitor and 5-FU at different concentrations on malignant melanoma and their antitumor mechanisms. It can provide experimental evidence for the clinical chemotherapy drug and improve adjuvant therapy of malignant melanoma after surgeryMethod:B16 melanoma cells were routinely cultivated in DMEM medium containing 10% new-born calf serum,0.1 g/L benzylpenicillin and 0.1 g/L xanthomycin.The study was divided into four experimental groups which was group A, group B, group C and group D. Group A was applied to only the ERK1/2 inhibitor, group B only 5-FU, group C combination of the ERK1/2 inhibitor and 5-FU and group D the control group.MTT assays was used to detect the inhibitory effects of proliferation on the B16 cells.Fow cytometry was used to detect the cell cycle and the rate of apoptosis.Take picture under microscope and reverse transcription-polymerase chain reaction was used to determine the mRNA levels of Bcl-2 and Caspase-9.Results:1 Combination of the ERK1/2 inhibitor and 5-FU showed a dose-dependent inbititio--n of B16 cell proliferation, compared to the effeccts of each agent alone and the control group(P<0.05).2 Combination of the ERK1/2 inhibitor and 5-FU showed a dose-dependent changes in cell phase, compared to the effeccts of each agent alone and the control group (p< 0.05).3 Microscopic observation showed that the control group were spindle, plump shape and only one drug had little change.The combination group had cell shrinkage, cracking, irregular shape, exfoliated cells increased and some cells appeared pseudo--podia at both ends of slender.This indicated the combination significantly inhibited the growth of B16 cells and cells showed significantly obvious morphological hanges.4 Combination of the ERK1/2 inhibitor and 5-FU potentiated the apoptosis and promoted the rate of apoptosis, compared to the effects of each agent alone and the control group(p<0.05).5 Combination of the ERK1/2 inhibitor and 5-FU potentiated a reduction in gene bcl-2 expression and upregulation of Caspase-9 expression.The difference was statistic-ally significant(P＜0.05).Conclusion:These results indicated that combination of the ERK1/2 inhibitor and 5-FU can inhibit the proliferation of B16 cells,accelerate apoptosis and join anti-cancer effect. Anti-cancer mechanism may be associated with activation of apoptosis, reduced gene bcl-2 expression and the upregulation of Caspase-9 expression.So the combination therapy of the inhibitor of ERK1/2 and 5-FU represent a novel anti-cancer strategy, which will reduce the toxicity and enhance tumor suppression.lt can provide experimental evidence for the clinical chemotherapy drug and improve adjuvant therapy of malignant melanoma after surgery.