| Objective:To study the anti-tumor effects of sodium citrate combined with arsenic trioxide on human gastric cancer cell in vitro and to explore its possible mechanism. Observe both combination to cancer of the gastric cell apoptosis is has cooperative after, for the clinical inhibition stomach cancer growth and to improve patient survival rate to provide basis ofexperimental.Methods:Treatment body rumored cultured gastric cancer cell line SGC-7901 (SGC) and BGC-803(BGC) with sodium citrate (final concentration was 5mmol/L) and(or)different concentrations of arsenic trioxide (final conce-ntration were 5,10umol/L).Cell proliferation was observe to by fluoresc-ent inversive microscopy.Trypan blue cellcount, them the cell apoptosis rate and phases of cell cycle were determined by flow cytometry. Observed expression of apop- tosis related genes of bcl-2 and mcl-1.The experimental datas classification.then use of Excel software to count. Measuring material expressed with mean differences±standard deviation, use SPSS 17.O software to statistics and analysis for data. Compares between Group and group use repetitive measure the variance analysis and single variable variance analysis. Compares of the one and one use SNK method. It is statistical significance in P< 0.05.Results:1.Compared with controls,add medicine group in the dosing intervention 24 hoursand 48 hours after, cells form observation:Living cell count and cell growth inhibition rate.Through the fluorescence microscope. The control group cell growth more than in the shape of the Angle,cells grow well stick wall, the density of growth rate and increased over time.The two plant cells proliferation are different degree of suppression,after 24hour show:Low concentration of two kinds of drug use, and two drugs low concentration of share, floating more cells, cells to contract go round to take off the wall floating, cell density are different degree of drop.After 48 hours, show above is more obvious.Compared with the control, the concentration of drug use group and two drugs share of the group, two plant cells live cell count all has different rate reduce. Cell proliferation inhibition rate than that of the control group has different rate increases,48hours with 24hours comparison, it is more obvious.2. The cell apoptosis①The apoptosis rate compared with the control group is increased (P<0.05).②The same cell, the same time, AS 10 umol/L group of apop-tosis rate was significantly higher than AS 5 umol/L group (P< 0.05).③The same cell, the same time, the share of the two drugs group cell apopt- osis rate was significantly higher than two drugs of the concentration of the corresponding use (P< 0.05).④The same time,CT 5 mmol/L share AS 10 umol/L group apoptosis rate is higher than CT 5 mmol/L or AS 5 umol/L group (P<0.05).⑤the same cell, the same drug concentration, 48 hours and 24 hours,also have statistical significance in one and one.3. The cell cycle changesTwo cells strains were treatment by the drug,through the flow cyto-metric detection,obviously been arrest at blockG2/M period, G2/M period cells up, G0/G1 phase significantly reduce, dosing group and blank compared with controls are statistically significant difference (P <0.05). The cell division is restr- ained, apoptosis rate increase. The control group did not see obvious the diploid peak, each group are detected the typical the diploid peak.4. Cell resistance apoptosis genes of expression bcl-2 and mcl-1①Compared with the control, the Bcl-2 and Mcl-1 gene expression which is relative quantity is down.②The same cell, the sametime, AS lOumol/L group of gene expression relatively lower than AS 5 umol/L group.③The same cell,the sametime,the share of two drugs of gene expression relative decline than alone use both of the two drugs the corre-sponding concentration group.④The same cell, the sametime, the CT 5 mmol/L share AS 10 umol/L group of genes relatively lower amount of expression CT5 mmol/L share AS 5umol/L group to decrease(P<0.05). The experimental results show that as the increase of the drug concentr-ation,gene expression relative reduction.Sodium citrate combining with arsenic trioxide had a stronger inhibition function on human gastric can- cer cell line than citrate or As2O3(P<0.05).Compared with blank group, the cell cycle distribution was altered and the ration of G0/G1 phase cell decreased and G2/M phase increased (P<0.05). After intervened by sod-ium citrate combining with arsenic trioxide the expression of bcl-2 and mcl-1 were down (P<0.05)Conclusion:Sodium citrate combining with arsenic trioxide has the synergistic effects on gastric cancer cell,and the mechanisms may be related to cell cycle blocking and apoptosis-inducing effects through regulating the expression of the related genes. |
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