论文部分内容阅读
目的:观察吉西他滨(gemcitabine,GEM)对卵巢癌移植瘤小鼠肿瘤免疫微环境的调节作用。方法:C57BL/6小鼠皮下注射卵巢癌ID8细胞构建卵巢癌移植瘤模型,实验组腹腔注射GEM,对照组注射生理盐水,观察肿瘤生长情况。流式细胞术检测小鼠脾脏及肿瘤组织的调节性T细胞(regulatory T cells,Treg)及髓来源抑制性细胞(myeloid-derived suppressor cell,MDSC),实时定量PCR检测小鼠肿瘤组织arginase-1(Arg-1)和Foxp3 mRNA的表达,流式细胞仪检测小鼠脾脏CD8+T细胞的比例,ELLSE法检测小鼠血清IFN-γ及IL-2的表达。结果:GEM处理后的移植瘤小鼠肿瘤生长明显受到抑制[(366.8±44.88)vs(499.3±24.14)mm~3,P<0.01]。移植瘤小鼠肿瘤组织及脾脏Treg比率明显降低[(12.71±2.31)%vs(20.36±2.65)%、(10.09±1.69)%vs(13.79±1.31)%,均P<0.01]。实验组的相关基因m RNA表达较对照组显著降低[Foxp3:(4.30±0.46)vs(6.35±0.58);Arg-1:(16.32±0.38)vs(13.26±0.37);均P<0.01]。实验组血清中的IFN-γ和IL-2含量明显高于对照组[IFN-γ:(71.90±2.28)vs(53.91±3.91)pg/ml;IL-2:(51.46±1.69)vs(40.90±1.50)pg/ml,均P<0.01]。结论:GEM下调卵巢癌移植瘤小鼠免疫抑制活性,并可上调小鼠抗肿瘤免疫原性,该结果可为GEM作为卵巢癌免疫治疗干预措施提供实验依据。
Objective: To observe the effect of gemcitabine (GEM) on tumor immune microenvironment in ovarian cancer xenografts in mice. Methods: C57BL / 6 mice were injected subcutaneously with ovarian cancer ID8 cells to construct ovarian cancer xenograft model. The experimental group was given intraperitoneal injection of GEM and the control group was injected with normal saline to observe the growth of the tumor. Flow cytometry was used to detect regulatory T cells (Tregs) and myeloid-derived suppressor cells (MDSCs) in spleen and tumor of mice. Real-time quantitative PCR was used to detect the expression of arginase-1 (Arg-1) and Foxp3 mRNA were detected by flow cytometry. The proportion of CD8 + T cells in spleen was detected by flow cytometry. The levels of IFN-γ and IL-2 in serum were detected by ELLSE. Results: The growth of tumor in GEM treated mice was significantly inhibited ([(366.8 ± 44.88) vs (499.3 ± 24.14) mm ~ 3, P <0.01]. The Treg rate of tumor and spleen in tumor-bearing mice was significantly lower than that in the control group ([(12.71 ± 2.31)% vs (20.36 ± 2.65)% vs (10.09 ± 1.69)% vs (13.79 ± 1.31)%, both P <0.01]. The expression of m RNA in the experimental group was significantly lower than that in the control group [Foxp3: (4.30 ± 0.46) vs (6.35 ± 0.58; Arg-1: (16.32 ± 0.38) vs (13.26 ± 0.37) vs P <0.01]. The levels of IFN-γ and IL-2 in the serum of the experimental group were significantly higher than that of the control group [IFN-γ: (71.90 ± 2.28) vs (53.91 ± 3.91) pg / ml; IL-2: ± 1.50) pg / ml, all P <0.01]. CONCLUSION: GEM can down-regulate the immunosuppressive activity of ovarian cancer xenografts in mice and up-regulate the anti-tumor immunogenicity of mice. This result may provide an experimental basis for GEM as an immunotherapy intervention for ovarian cancer.