| Part oneObjective:To observe the therapeutic effect of 5-HT combined with PD-1 inhibitor on the treatment of mice with colon cancer xenograft.Methods:Colon cancer MC38 cells were inoculated into the right hypochondrium of C57BL/6 mice.When the subcutaneous tumour volume reached100-150mm3,the mice with successful subcutaneous tumour implantation were randomly divided into 6 groups: control group,5-HT(0.4mg/kg)group,5-HT(0.8mg/kg)group,PD-1 inhibitor group(250ug/mice),5-HT(0.4mg/kg)+PD-1 inhibitor group and 5-HT(0.8mg/kg)+PD-1 inhibitor group.The mice were anesthetized and executed at 25 d.Subcutaneous tumours were collected and measured from each group of mice and statistically analyzed.Results:1.The tumour volumes of the control group,5-HT(0.4mg/kg)group and5-HT(0.8mg/kg)group were similar and no statistical differences were observed(P>0.05).2.Compared with the control group,PD-1 inhibitor group,5-HT(0.4mg/kg)+PD-1 inhibitor group and 5-HT(0.8mg/kg)+PD-1 inhibitor group all significantly inhibited the growth of subcutaneous tumours(266.7±386.5mm3 vs.1675.0±838.2mm3,P<0.01;11.0±10.3mm3 vs.1675.0±838.2mm3,P < 0.01;525.0±50.0mm3 vs.1675.0±838.2mm3,P <0.01).3.The tumour volume of the 5-HT(0.4mg/kg)+PD-1 inhibitor group was significantly smaller than that of the PD-1 inhibitor group(11±10.3mm3 vs.266.7±386.5mm3,P < 0.05).The tumour volume of the 5-HT(0.8mg/kg)+PD-1 inhibitor group was larger than that of the PD-1 inhibitor group,but no statistical difference was observed(P>0.05).Conclusions:1.0.4mg/kg 5-HT enhanced the tumour suppressive effect of PD-1inhibitor on colon tumour-bearing mice;2.The application of 5-HT alone did not promote or inhibit the growth of colon cancer transplanted tumours in mice.Part twoObjective:To analysis of 5-HTR mRNA expression on CD8+T cells and HCT116 cells.Methods:Peripheral blood mononuclear cells from healthy volunteers were extracted.CD8+T cells were isolated and purified using the immunobead method,and mouse anti-human CD3 and CD28 monoclonal antibodies were applied to activate CD8+T cells.HCT116 cells were recovered,cultured and passaged.Total RNA from activated CD8+T cells and HCT116 cells were extracted and reverse transcribed into c DNA.q RT-PCR was applied to comparatively analyze the mRNA expression of 5-HTR on CD8+T cells and HCT116 cells.Results:The purity of CD8+T cells reached 99.4% and the activation level was96.5%.The mRNA expression of 5-HTR2 B on activated CD8+T cells was significantly higher than that of HCT116 cells(P<0.01),while the mRNA expression of 5-HTR1 A,5-HTR1 B,5-HTR2 C and 5-HTR7 were significantly lower than those of HCT116 cells(P<0.05).The mRNA expression of5-HTR2 A,5-HTR3 A and 5-HTR3 B on activated CD8+T cells and HCT116 cells showed no statistical difference(P>0.05).Conclusions:The mRNA expression of 5-HTR differed between activated CD8+T cells and HCT116 cells;The mRNA expression of 5-HTR2 B on activated CD8+T cells was significantly higher than that of HCT116 cells;The mRNA expression of 5-HTR1 A,5-HTR1 B,5-HTR2 C and 5-HTR7 on activated CD8+T cells were significantly lower than that of HCT116 cells. |
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