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目的:探讨骨髓间充质干细胞(Mesenchymal Stem cells,MSCs)的全细胞抗原(WCAs)对于人肺腺癌细胞株A549移植瘤的预防接种作用,为发掘肿瘤免疫治疗提供新策略。方法:采用全骨髓贴壁法原代培养小鼠MSCs并以流式细胞仪鉴定,取3~5代MSCs细胞以15Gy X线灭活获取(Whole cell antigens,WCAs),以该抗原皮下接种于BALB/c小鼠(1次/3 d,共2周),获得免疫接种小鼠模型,对照组皮下注射同体积的PBS;为了方便示踪,以Lipofectamine TM 2000细胞脂质体转染绿色荧光蛋白(GFP),获得标记GFP-A549细胞株,皮下移植瘤株进行荷瘤,采用小动物荧光成像仪观察肿瘤生长;同时评估肿瘤直径并计算肿瘤体积,行Ki-67免疫组化染色初步分析肿瘤增殖能力。结果:肺腺癌GFP-A549细胞株皮下移植成功使小鼠荷瘤,实验组小鼠的肿瘤体积显著小于对照组(★P<0.05,★★P<0.01),小动物荧光成像仪结果与解剖结果一致;实验组Ki-67的表达低于对照组。结论:本研究采用MSCs获得WCAs在荷瘤前对小鼠进行免疫刺激,发现其确实有抑制肿瘤生长的作用。
Objective: To investigate the vaccination effect of all-cell antigen (WCAs) of mesenchymal stem cells (MSCs) on human lung adenocarcinoma A549 xenografts and to provide a new strategy to explore tumor immunotherapy. Methods: MSCs were primary cultured with whole bone marrow adherent method and identified by flow cytometry. Whole-cell antigens (WCAs) were harvested from 3 to 5 passages of MSCs and subcutaneously inoculated with BALB / c mice (1 time / 3 days for 2 weeks) were immunized with mouse model. The control group was injected subcutaneously with the same volume of PBS. To facilitate the tracing, Lipofectamine ™ 2000 cells were transfected with green fluorescence (GFP), get labeled GFP-A549 cell line, subcutaneously transplanted tumor-bearing tumor bearing small animal fluorescence imaging instrument observed tumor growth; at the same time assess the tumor diameter and calculate the tumor volume, line Ki-67 immunohistochemical staining Tumor proliferation ability. Results: Tumor bearing mice were successfully transplanted into the lung adenocarcinoma cell line GFP-A549 successfully. The tumor volume of the mice in the experimental group was significantly smaller than that of the control group (P <0.05, P <0.01, P <0.01) The anatomical results were consistent. The expression of Ki-67 in the experimental group was lower than that in the control group. Conclusion: In this study, MSCs were used to obtain WCAs before immunostimulation in mice and found that they did inhibit tumor growth.