| ObjectiveTo observe the influence of Haishengsu on transplanted tumor of nude mice inoculated with K562 and K562/AO2 cells,and try to find out the mechanism.MethodsInoculated BALB/c nude mice subcutaneously with K562/AO2 and K562 cells, respectively. When the transplanted tumor came up, injected intravenously different dose of Haishengsu and set up a combination therapy group, blank and positive control group. And then measured the changes in tumor size regularly to draw the tumor growth curve ,calculated the relative tumor volume(RTV), tumor inhibitory rate (IR%); pathology observed; FCM to detect the apoptosis and cell cycle of the tumor tissue,also to detect the express of p-gp.Results1.Tumor Inhibitory Rate: Group of the K562 cells: inhibitory rate of adriamycin group, the combination therapy group(ADR+HSS), HSS-high, HSS-medium and HSS-low group were 88.91%, 85.81%, 99.96%, 96.41%, 53.41%, respectively.Effects of HSS-medium group(900mg/kg) and HSS-high group(1800mg/ kg)was better than doxorubicin group and the combination group(p <0.05), effect of HSS-low group was worse than other groups(p<0.01); K562/AO2 cell:inhibitory rate of adriamycin group, the combination therapy group(ADR+HSS), HSS-high, HSS- medium and HSS-low group were 23.07%, 52.86%, 99.96%, 95.89%, 44.13%, respectively. Effects of HSS-medium group(900mg/kg)and HSS-high group(1800mg/ kg)was better than doxorubicin group and the combination group(p<0.01), low-dose group less effect.2.Pathology of tumor: There were a large number of tumor cells in the control group(PBS), much nuclear fission; K562-HSS group: angiogenesis, necrosis and bleeding could be easily seen, and much inflammatory cells,much necrosis of cells, the nuclear enrichment and broken Crack could be seen easily, we can see some of the apoptotic bodies; K562/AO2-HSS group shows that a large number of tumor cells necrosis, the cells array in disorder, structure not clear, the nuclear enrichment and broken Crack could be seen easily.3.Apoptosis and Cell Cycle: K562 cell groups: the early apoptosis rate of low-dose group of HSS was 0.01%, late apoptosis rate accounted for 0.91%; the late apoptosis rate of middle dose group of HSS was 0.93%; the late apoptosis rate of high dose group of HSS was 80.88%.K562/AO2 cell groups: the early apoptosis rate of low-dose group of HSS was 0.02%, late apoptosis rate accounted for 0.88%; the early apoptosis rate of middle dose group of HSS was 0.06%, late apoptosis rate accounted for 5.20%; the early apoptosis rate of high dose group of HSS was 0.01%, late apoptosis accounted for 87.49%. With the dose increased, the proportion of apoptosis increased significantly, HSS of high-dose group of HSS induced apoptosis mainly in late phase. HSS do not effect the cell cycle in Group of the K562 cells,but can block the cell cycle in phase of G0-G1 in Group of the K562/AO2 cells.4.The expression of p-gp: K562 cells groups: Expression of p-gp protein range in 1.86-3.53%, and low protein expression.K562/AO2 cell groups: p-gp protein expression of the low, middle and high dose group rates were 95.11%, 92.07%, 86.68%,respectively.with the dose of HSS increasing,the p-gp protein expression gradually decreased, especially the high-dose group.5.RT-PCR for the expression of mdr1:K562 cells groups: without the expression of mdr1 gene.K562/AO2 cells groups: With the dose of HSS increasing, mdr1 gene expression reduced gradually.6.RT-PCR for the expression of BCR/ABL:K562 cells groups: without the expression of BCR/ABL gene.K562/AO2 cells groups: With the dose of HSS increasing, BCR/ABL gene expression reduced gradually.7.RT-PCR for the expression of sorcin:K562 cells groups: without the expression of sorcin gene.K562/AO2 cells groups: With the dose of HSS increasing, sorcin gene expression reduced gradually. Conclusion:Haishengsu could inhibit the growth of transplanted tumor of nude mice significantly, and can reverse resistance of transplanted tumor(K562/AO2) to Adriamycin.
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