Effect Of Sorafenib Combined With Salinomycin On Proliferation Of Human Gastric Cancer MGC-803 Cells And Regulation Of The RAF/MEK/ERK Pathway

Objective: To observe the effect of sorafenib and salinomycin on the proliferation of human gastric cancer cell line MGC-803 in vitro,And to explore the effect of sorafenib on the intracellular RAF / MEK / ERK signaling pathway and possible molecular mechanisms.Methods: Human gastric cancer MGC-803 cell line as the research object,MGC-803 cells were treated with sorafenib and salinomycin at different concentrations(0.375μg / m L,1.5μg / m L,6μg / m L,24μg / m L and 96μg / m L)for 24 hours,48 hours,72 hours.Cell proliferation was detected by MTT assay.MGC-803 cells were treated with sorafenib and salinomycin at a concentration of 6μg / m L for 48 h,Cell proliferation was detected by MTT assay.Inverted phase contrast microscope was used to observe the morphological changes of cells.Western blot detection of p ERK and ERK protein expression.Results: The inhibitory rates of sorafenib on MGC-803 cells at a certain concentration range(0.375μg / m L,1.5μg / m L,6μg / m L,24μg / m L and 96μg / m L)for 24 hours,48 hours and 72 hours were 5.37%、12.71%、23.38%、44.54%、78.38%;13.52%、24.64%、43.53%、62.47%、89.17%;16.28%、38.53%、67.54%、71.15%、97.19%.The inhibitory rates of Salinomycin to MGC-803 cells at a certain concentration(0.375μg / m L,1.5μg / m L,6μg / m L,24μg / m L and 96μg / m L)for 24 h,48h and 72 h were 5.93%、10.37%、26.96%、39.36%、78.58%;10.16%、20.70%、40.36%、79.16%、89.19%;15.49%、39.68%、56.32%、88.73%、99.2%.The results showed that MGC-803 cells treated with sorafenib and salinomycin alone or in combination could inhibit cell proliferation in different concentrations.Inverted phase contrast microscope showed: Negative control group cells grew well;MGC-803 cells were treated with sorafenib or salinomycin alone for 48 hours,A typical form of apoptosis were observed under the microscope,Combined treatment group more obvious apoptosis.Western blot results showed: More negative control group and salinomycin group,The sorafenib group and combination group ERK protein expressions were no significant difference;Compared with the negative control group and salinomycin group,the sorafenib group and the combination group obviously down-regulated the expression of p ERK protein,and the down-regulation of the combination group was more obvious.Conclusions:(1)Sorafenib and salinomycin can effectively inhibit the proliferation of human gastric cancer MGC-803 cells in a dose-dependent and time-dependent manner.(2)The effect of combination therapy is stronger than that of single drug.The combined use of drugs has a synergistic effect on inhibiting proliferation of human gastric cancer MGC-803 cells in vitro.(3)Sorafenib Regulates the RAF/MEK/ERK Signaling by Downregulating p ERK Protein Expression.