CAJ | 학술논문

目的：研究FOLFOX4与1-D-甲基色氨酸(1-D-methyl tryptophan，1-D-MT)联合应用是否有利于改善荷胃癌小鼠的免疫耐受状态。方法：用脂质体转染法，将pcDNA3.1-IDO质粒和pcDNA3.1(+)空质粒稳定转染MFC细胞，并设未转染组；用RT-PCR和Western blot法检测未转染组、空质粒转染组和pcDNA3.1-IDO转染组IDO mRNA和蛋白的表达；然后建立荷胃癌小鼠皮下移植瘤模型，并设未转染组、空质粒转染组，IDO+生理盐水(NS)组、FOLFOX4组、1-D-MT组和FOLFOX4+1-D-MT联合处理组，采用流式细胞术检测各组小鼠脾脏中Treg细胞数量变化；RT-PCR检测FOXP3 mRNA表达量变化。结果：IDO mRNA与蛋白表达在稳定转染pcDNA3.1-IDO质粒组细胞较未转染组、空质粒转染组表达量明显增加(P<0.05)。IDO+NS组小鼠脾脏Treg细胞比例及FOXP3 mRNA含量较未转染组、空质粒组均明显增多(P<0.05)。FOLFOX4+1-D-MT组和1-D-MT组与FOLFOX4组相比，Treg细胞比例及FOXP3 mRNA含量均明显降低(P<0.05)，且联合治疗组较1-D-MT组降低更明显(P<0.01)。结论：通过抑制Treg的增殖及降低FOXP3 mRNA表达，FOLFOX4与1-D-MT联合应用可降低机体免疫逃逸能力。
목적：연구FOLFOX4여1-D-갑기색안산(1-D-methyl tryptophan，1-D-MT)연합응용시부유리우개선하위암소서적면역내수상태。방법：용지질체전염법，장pcDNA3.1-IDO질립화pcDNA3.1(+)공질립은정전염MFC세포，병설미전염조；용RT-PCR화Western blot법검측미전염조、공질립전염조화pcDNA3.1-IDO전염조IDO mRNA화단백적표체；연후건립하위암소서피하이식류모형，병설미전염조、공질립전염조，IDO+생리염수(NS)조、FOLFOX4조、1-D-MT조화FOLFOX4+1-D-MT연합처리조，채용류식세포술검측각조소서비장중Treg세포수량변화；RT-PCR검측FOXP3 mRNA표체량변화。결과：IDO mRNA여단백표체재은정전염pcDNA3.1-IDO질립조세포교미전염조、공질립전염조표체량명현증가(P<0.05)。IDO+NS조소서비장Treg세포비례급FOXP3 mRNA함량교미전염조、공질립조균명현증다(P<0.05)。FOLFOX4+1-D-MT조화1-D-MT조여FOLFOX4조상비，Treg세포비례급FOXP3 mRNA함량균명현강저(P<0.05)，차연합치료조교1-D-MT조강저경명현(P<0.01)。결론：통과억제Treg적증식급강저FOXP3 mRNA표체，FOLFOX4여1-D-MT연합응용가강저궤체면역도일능력。
OBJECTIVE: To investigate whether combining 1-D-MT with FOLFOX4 could be useful to TMimprove the immune tolerance of gastric cancer-bearing mice. METHODS:By using the lipofectamine 2000,the eukaryotic expression plasmid pcDNA3.1-IDO and empty vector pcDNA3.1 (+) were transfected in a MFC cell line,setting a control group. The expression of IDO was detected by reverse transcription polymerase chain reaction(RT-PCR) and western blot. Animal model of gastric cancer-bearing mice were established to receive normal saline (NS),FOLFOX4,1-D-MT and 1-D-MT+FOLFOX4 therapy. By using flow cytometry,Treg cell ratio change was analyzed and FOXP3 mRNA expression change was assessed by RT-PCR. RESULTS:The expression of IDO mRNA and protein in the group trancfected with pcDNA3.1-IDO was obviousely ligher than the MFC group and the empty vector group (P<0.05). Treg cell ratio and FOXP3 mRNA expression in the transfected IDO negative control group with NS therapy was higher than the MFC and the empty vector groups (P<0.05). The Treg ratio and FOXP3 mRNA in 1-D-MT+FOLFOX4 and 1-D-MT groups were less than in FOLFOX4 group(P<0.05),with 1-D-MT+FOLFOX4 group more markedly than 1-D-MT group (P<0.05). CONCLUSION:Combining 1-D-MT with FOLFOX4 could reduce Treg cell ratio and FOXP3 expression,thus reducing the immune escape.