| Objective: Multiple myeloma (MM) is a disease of malignant and anti-apoptosised of plasma cell. At present, MM should not be completely cured, the mainly treatment is chemotherapy, the patient's 5-year survival rate is less than 25%. Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), also known as Apo2Lligand, belongs to one member of TNF superfamily. TRAIL can induce cells apoptosis and the majority of normal cells had no effect on. Recombinant mutant human TRAIL (rmhTRAIL), is increased of TRAIL's stability, solubility and biological activity. At the early stage of myeloma research, it has been found with the multi-drug rmhTRAIL united, the myeloma cells can enhance cell-surface death receptor expression, reduce expression of tumor suppressor genes. To further, verifying the anti-tumor effect of rmhTRAIL in vivo, we use models of RPMI-8226 cells transplanted in nude mice, observe the effect of anti-tumor and drug toxicity in vivo with rmhTRAIL united dexamethasone, thalidomide, provide basis for clinical application.Methods:Transplant tumor in nude mice: BALB / C-nu / nu nude mice are reared under SPF conditions, taking the PRMI-8226 cells to inoculate into the right shoulder back of nude mice. After the passage., remove the transplanted tumor then cut into 70 ~ 100mm3 ,inoculate into right shoulder back of nude mice, at the same time, some tumor cells labeled homogenate to observe its morphology. These tumor volume grow to 300-500mm3 size, check without spontaneous hemorrhage and necrosis of tumor-bearing nude mice infected as subjects:1. grouping and delivery: Based on the size of these tumor, the mice were divided into 6 groups: GroupA (non-drug treatment group), groupB (rmhTRAIL monotherapy group), rmhTRAIL 5mg/kg d1-10; groupC (dexamethasone monotherapy group), dexamethasone 8mg/kg d1-4 ,9-12; groupD (thalidomide monotherapy group), thalidomide 75mg/kg d1-10; groupE (rmhTRAIL United dexamethasone group), rmhTRAIL 5mg/kg d1-10, dexamethasone 8mg/kg d1-4 ,9-12; groupF (rmhTRAIL United thalidomide group), rmhTRA IL 5mg/kg d1-10, thalidomide 75mg/kg d1-10. Thalidomide used the administration of gavage, the others of intraperitoneal injection. After d20 sacrifice the survival of nude mice(1) rmhTRAIL in combination with dexamethasone observe the groups of A, B, C, E.(2) rmhTRAIL in combination with thalidomide observe the groups of A, B, D, F.2. To assess the antitumor activity: every other day before delivery measure tumor's length (a) and short diameter (b), based on the formula, tumor volume =ab2/2, the relative size = tumor volume after administration/tumor volume before administration, calculate the tumor size and relative size. Until the end of all treatment, according to the formula: inhibitory rate =(1-the average tumor size of the treated group /the average tumor volume of the control group )×100%, to calculate the inhibition rate of the treatment group.3. Observed the adverse effects of drugs in nude mice, including the mental state of tumor-bearing mice, the number of water consumption, body weight changes.4. Histopathological examination: D20 executed all the nude mice, The specimens of transplantation tumors were stained with HE, observe the tumor tissue under the intensity of tumor cells (each 40 times the field of vision) and cell morphology.5. The results of experiment were analyzed by SPSS10.0.The different of the volume of the transplantation tumor before or afte radministration in eaeh group of the bearing nude mice were test by repeated measure ANOVA. The differences of relative tumor volume bearing nude mice between four experimental groups were test by one-way ANOVA .Ps of less than 0.05 were considered significant.Results:1. Transplanted tumor cells morphology: transplanted tumor cells homogenate, almost all of the cells are original plasma cells, some of multinucleated myeloma cells are in split-phase.2. In accordance with tumor volume analysis the proliferation inhibitory effect of three kinds of drugs on the RPMI-8226 cell: The rate of tumor volume increase in groupA was the fastest,followe by groupB groupC and groupD. GroupA the volume from the D0 (373.5±40.0) to D20 (12824.5±6960.7), difference is significant (P <0.05); group B transplanted tumor volume decrease from D0 (394.8±78.1) reduced to D20 (12.0±18.8), differences have statistical significance (P <0.05); group C (dexamethasone monotherapy group) must have anti-tumor effect, but no significant inhibitory effect , transplanted tumor size increased gradually from D0 (394.8±78.1) to D20 ( 4754.8±2115.2) (P <0.05), D20 was still less than the group A (P <0.05); group D, simply thalidomide showe no obvious inhibitory effect , transplanted tumor volume increased gradually from D0 (411.1±78.2) increased to D20 (8178.7±6691.0) (P <0.05), D20 is also smaller than the A group; groupE at the beginning the tumor volume decreased from D0 (441.2±66.7) reduced to D12 (123.7±115.8 ), after all administered the tumor size increased rapidly to D20 (702.2±908.9), the observation time points have no significant difference (P>0.05), compare to groupB D20 tumor volume ( 12.0±18.8) < groupE (702.2±908.9) significantly (P <0.05), that is, from the analysis of terminal tumor volume ,rmhTRAIL United dexamethasone compared with the rmhTRAIL alone has no advantages; groupF RPMI-8226 cells significantly inhibited, transplanted tumor volume decrease from D0 (425.7±63.3) to D20 (0.0±0.0) (P <0.05), compared with the groupB, groupF tumor size of D20 had no significant difference (P>0.05), purely from the terminal tumor volume, the effect of rmhTRAIL united is not better than the rmhTRAIL alone.3. In accordance with relative tumor volume analysis the proliferation inhibitory velocity of three kinds of drugs on the RPMI-8226 cell: (1) rmhTRAIL in combined with dexamethasone: comparison with the groupA, at D2 nude relative volume of groupC has no significant difference ( P=0.075), beginning from D4 relative volume of groupC is relatively smaller than groupA, suggest that dexamethasone has dose cumulative, tumor volume grow slowly; the group which contain rmhTRAIL including groupB, groupE the relative volume is less than groupA from D2 ,has statistical significance (P <0.05); tumor relative volume of groupB were significantly less than the groupC at various observation points (P <0.05), groupB of the relative tumor volume is always smaller than the groupE. from the transplanted tumor growth inhibition rate of the volume suggust, GroupA < GroupC < GroupE < groupB ,rmhTRAIL single is the most effective drug group. (2) rmhTRAIL United thalidomide: comparison with the groupA, relative tumor volume of groupD there was no significant difference at different time points observation (P> 0.05); the group which contain rmhTRAIL including groupB, groupF the relative volume is less than groupA from D2 ,has statistical significance (P <0.05);compared with groupD the relative volume of groupB is less than groupD (P<0.05); groupB and group F ,beginning from D8 groupF (0.04±0.07) < groupB (0.13±0.14) to D20 F Unit Transplantation Relative tumor volume is always smaller than the groupB. The above tips from the transplanted tumor volume growth rate, GroupA |
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