| 【Objectives】1. Observe the proliferation inhibition and the apoptosis of human gastric cancer cellLine SGC-7901induced by Cinobufacini, arsenic trioxide and the combination of the twodrugs.2. Discuss if there is a synergistic effect between Cinobufacini and arsenic trioxide onhuman gastric carcinoma cell line SGC-7901.3. Explore new methods in the treatment of gastric cancer, provide some experimentalbasis for the clinical medication.【Methods】1.The human gastric cancer cell line SGC-7901cultured in vitro and in logarithmicgrowth phase were divided into negative control group, Cinobufacini (Cino) group, As2O3group and Cinobufacini+As2O3group. Then respectively use different concentrationsgradient of As2O3(1μmol/L、5μmol/L、10μmol/L) and Cinobufacini (0.125μg/ml,0.25μg/ml,0.5μg/ml) alone or combination of the two drugs on the gastric cancer SGC-7901cells.2.Using MTT (Methylthiazolyldiphenyl-tetrazolium bromide) reduction method todetect respectively the effect of Cinobufacini and As2O3, and to detect the combination ofthe two drugs on the growth of gastric cancer SGC-7901cells.3.Using an inverted phase contrast microscope to respectively observe thecell morphology of apoptosis on the negative control group, single drug-using group,combination of the two drugs group in24h,48h,72h. 4.Using flow cytometry (FCM) to detect respectively the apoptosis effect and the cellcycle of gastric cancer SGC-7901cells generated by Cinobufacini, As2O3, and thecombination of the two-drugs.【Results】1.Observe the cell morphology under inverted microscope, with the lowconcentration of Cinobufacini and As2O3, morphological changes of apoptosis in humangastric cancer SGC-7901cells could be seen, cells shrinked and turned round, theattachment of cells was poor, with high concentration of these two drugs, morphologicalchanges of cells characterized by necrosis, vacuoles in cells and a large number ofnecrotic debris were visible.2. Using MTT to make analysis, it showed that both Cinobufacini and As2O3had theeffect of proliferation inhibition in human gastric cancer SGC-7901cells. And with theconcentration of drug increased or the action time prolonged, the effect of proliferationinhibition was enhanced, which means that there was a positive correlation betweendrug’s concentration and action time. Compared with the groups treated just by one drug,the difference of cells’ proliferation inhibition between the combination groups and onedrug action groups had statistically significant (P<0.05).3. Cell cycle analysis showed that, compared with the control group, when thehuman gastric cancer SGC-7901cells affected by Cinobufacini, the percentage of G0/G1phase cells increased, the proportion of cells in S phase decreased, cells were arrested inG0/G1phase. Whereas when the human gastric cancer SGC-7901cells affected byAs2O3, the percentage of S phase cells increased, the proportion of cells in G0/G1phasereduced, cells were arrested in G0/G1phase. After the human gastric cancer SGC-7901cells affected by the combination of two drugs for48h, the percentage of cells both inG0/G1phase and S phase increased, cells were arrested in G0/G1and S phase(P<0.05).4. Both Cinobufacini and As2O3had the effect of apoptosis induction on the humangastric cancer SGC-7901cells. Compared with the groups treated by Cinobufacini orAs2O3, the difference of the apoptosis rate of human gastric cancer SGC-7901cells between the combination groups and one drug action groups had statistically significant(P<0.05).【Conclusions】Both Cinobufacini and arsenic trioxide could inhibit the growth of human gastriccancer SGC-7901cells. Both of them had proliferation inhibiting and apoptosis inducingeffect on gastric cancer SGC-7901cells. The combination of the two drugs could play asynergistic effect on gastric cancer cells in inhibiting proliferation and inducing apoptosis,and this kind of killing effect was positively related to the drugs’ concentration and actiontime. |
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