白玉菇多糖对宫颈癌荷瘤小鼠肿瘤生长和细胞因子的 影响及机制

Effect of white shimeji mushroom polysaccharides on tumor growth and cytokines in mice with cervical cancer and its mechanism

目的探讨白玉菇多糖对宫颈癌荷瘤小鼠肿瘤生长及血清细胞因子的影响。方法50只BALB/c雌性小鼠通过皮下接种小鼠宫颈癌U14细胞株建立宫颈癌荷瘤小鼠模型,将建模成功小鼠随机分为模型组、胸腺肽组、白玉菇多糖组、FGFR3抑制剂组及白玉菇多糖+FGFR3抑制剂组;末次给药24 h后观察记录小鼠瘤体积和瘤质量,并计算免疫器官指数;流式细胞术检测外周血T淋巴细胞亚群(CD4^(+) T、CD8^(+) T)比例和CD4^(+) /CD8^(+) 比值,酶联免疫吸附剂(ELISA)检测肿瘤坏死因子-α(TNF-α)、白细胞介素-2(IL-2)、转化生长因子-β(TGF-β)及白细胞介素-10(IL-10)水平,实时荧光定量聚合酶链反应(RT-qPCR)和蛋白质免疫印迹法(WB)检测肿瘤组织中FGFR3、血管内皮生长因子(VEGF)、程序性死亡因子配体1(PD-L1)、细胞增殖指数(Ki-67)信使核糖核酸(mRNA)及蛋白表达。结果与模型组比较,胸腺肽组、FGFR3抑制剂组、白玉菇多糖组和白玉菇多糖+FGFR3抑制剂组小鼠瘤体积、瘤质量、CD8^(+) T淋巴细胞比例及TGF-β、IL-10水平降低,抑瘤率、胸腺和脾指数、CD4^(+) T淋巴细胞比例、CD4^(+) /CD8^(+) 比值及TNF-α、IL-2水平升高(P<0.05),小鼠肿瘤组织FGFR3、VEGF、PD-L1、Ki-67 mRNA及蛋白表达降低(P<0.05);与胸腺肽组、FGFR3抑制剂组和白玉菇多糖组比较,白玉菇多糖+FGFR3抑制剂组对上述指标的作用更加显著(P<0.05)。结论白玉菇多糖对宫颈癌荷瘤小鼠肿瘤生长有显著抑制作用,其机制可能是通过抑制FGFR3及下游VEGF、PD-L1的表达,提高机体免疫力而实现的。

Objective To investigate the effect and mechanism of white shimeji mushroom polysaccharides on tumor growth and cytokines of mice with cervical cancer.Methods Fifty mice were subcutaneously inoculated with mouse cervical cancer U14 cell line to establish a cervical cancer tumor-bearing mouse models.The successfully modeled mice were randomly divided into the model group,the thymosin group,the white shimeji mushroom polysaccharides group(shimeji group),FGFR3 inhibitor group and the white shimeji mushroom polysaccharides+FGFR3 inhibitor group(shimeji+FGFR3 group).The tumor volume and mass were observed and recorded 24 h after the last administration,and the immune organ index was calculated.The ratio of peripheral blood T lymphocyte subsets(CD4^(+) T,CD8^(+) T)and the ratio of CD4^(+) /CD8^(+) were detected by flow cytometry.The levels of tumor necrosis factor-α(TNF-α),interleukin-2(IL-2),transforming growth factor-β(TGF-β),and interleukin-10(IL-10)were determined by enzyme-linked immunosorbent(ELISA).The messenger ribonucleic acid(mRNA)and protein expressions of FGFR3,vascular endothelial growth factor(VEGF),programmed cell death ligand 1(PD-L1),cell multiplication index(Ki-67)in tumor tissues were detected by real-time quantitative polymerase chain reaction(RT-qPCR)and western blotting(WB)respectively.Results Compared with the model group,the tumor volume,tumor mass,CD8^(+) T lymphocyte ratio,TGF-β,and IL-10 levels significantly decreased in the thymosin group,shimeji group,FGFR3 inhibitor group and Shimeji+FGFR3 group(P<0.05),while tumor inhibition rate,thymus and spleen index,CD4^(+) T lymphocyte ratio,CD4^(+) /CD8^(+) ratio,TNF-αand IL-2 levels increased significantly in those groups(P<0.05).The mRNA and protein expressions of FGFR3,VEGF,PD-L1,Ki-67 in tumor tissues of the thymosin group,shimeji group,the FGFR3 inhibitor group and shimeji+FGFR3 group significantly decreased(P<0.05).The treatment in shimeji+FGFR3 group had the strongest effect(P<0.05).Conclusion white Shimeji mushroom polysaccharides can significantly inhibit the tumor growth of cervical cancer bearing mice,which is speculated to be achieved by inhibiting the expression of FGFR3 and downstream VEGF,and PD-L1,and improving the immunity of the body.