| Objective:This experiment chooses SGC7901and SGC7901/VCR as the research object. Byadopting the method of serum pharmacology、CCK8, Western Blot, RT-PCR to graduatestudent vein injection of human gastric cancer SGC7901/VCR cells multi-drug resistancereversal agents, And explore its action mechanism and the related signal pathways. For theuse of chemotherapy with shengmai injection therapy in gastric cancer provideexperimental basis.Methods:1．To prepare the medicated serum of shengmai injection.Conventional cultivationof human gastric cancer cells SGC7901and the resistant cells SGC7901/VCR.2．Identification of SGC7901/VCR multi-drug resistance: CCK8test the IC50alu eof VCR, DDP, ADM,5-FU four chemotherapy drugs on SGC7901and SGC7901/VCRcell. Western Blot test and verify multidrug resistance and the resistance mechanis ms ofSGC7901/VCR. Using the flow cytometry to detect the cell cycle distribution char acter ofcells SGC7901and the resistant cells SGC7901/VCR.3．To test the effects of shengmai injection of serum contains on cells SGC7901andthe resistant cells SGC7901/VCR; CCK8detect toxic effects on cells of shengmaiinjection of serum containg and calculated dose toxicity of the drug-contains serum.4．Test the effects of shengmai injection of serum containg on Reversing multi-drugresistance: CCK8detect minimum toxic dose of drug-containing serum joint combination of four different chemotherapy drugs on the resistance cells, and calculate the IC50valuesand reversal of drug resistance index. Using Flow cytometry to detect the cell cycledistribution character of resistant cells before and after treatment with serum drug.5．To explore the Reversal of drug resistance mechanisms of shengmai injection ofserum contains：Western Blot detect The influence on pulse injection drug-containingserum on P-gp, c-jun and phosphorylation of c-jun expression in JNK signaling pathwayin resistant cells.RT-PCR detect The influence of drug-containing serum of shengmaiinjection on MDR1, MRP1expression in gastric cancer resistant cells.Results:1．The cytotoxic effect, drug-containing serum on SGC7901/VCR cell andSGC7901, Was positively correlated with serum concentrations. When the drug containting serum concentration of40%, The cell survival rate was91%.2．Compared with SGC7901cells,p-gp protein is highly expressed in SGC7901/VCR cells.c-jun and phosphorylated c-jun in JNK signaling pathway express InSGC7901/VCR less than SGC7901.JNK signaling pathway may be involved indrug-resistant.3．SGC7901/VCR cell resistant against VCR, ADM, DDP,5-FU and the IC50values are Higher than SGC7901.4．After treating by serum combining chemotherapy drugs on the drug resistantcells,IC50value lower than before.5．Cell cycle analysis showed that serum Containing shengmai can interfere withcell cycle,and Inhibit proliferation of resistant cells.6．Western Blot analysis showed that after drug treatment by shengmai serum-containing cells,levels of p-c-jun was higher than before.7．RT-PCR analysis showed that MDR1mRNA, MRP1levels in SGC7901/VCR were significantly higher than SGC7901，and after treating by serum Containingshengmai,MDR1mRNA, MRP1levels significantly decreased than before.Conclusion:1．Compared with SGC7901,SGC7901/VCR was resistance to chemo therapy drugs.2． Drug-containing serum of Sheng Mai injection can reversed the Resistan ce ofSGC7901/VCR.3． The effect of serum containing Shengmai may be achieved by increasing thephosphorylation of c-jun levels, inhibition of JNK signaling pathway, reducing MDR/P-gp levels... |
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