树突状细胞与黑色素瘤细胞体外共培养体系对小鼠黑色素瘤形成的影响

发布时间：2018-03-13 23:41

本文选题：黑色素瘤　切入点：树突状细胞　出处：《山东医药》2017年05期 　论文类型：期刊论文

【摘要】：目的探讨树突状细胞(DC)与黑色素瘤细胞(B16)体外共培养后对小鼠黑色素瘤形成的影响。方法提取小鼠骨髓细胞,分化DC,培养至第6天,将细胞分为4组,脂多糖(LPS)组、聚肌胞[Poly(IC)]组、Melan-A抗原肽(Melan-A)组分别加入DC佐剂LPS(终浓度100 ng/m L)、Poly(IC)(终浓度20 ng/m L)、Melan-A抗原肽(终浓度5μmol/L)进行处理,未处理组未进行干预。采用流式细胞术检测DC成熟状态。B16细胞与培养第6天的DC共培养2 d。取C57BL/6J小鼠皮下注射B16细胞(8只),DC+B16细胞(8只),LPS(8只)、poly(IC)(8只)、Melan-A抗原肽激活(8只)的DC+B16细胞,接种B16细胞1周再接种DC+B16细胞(5只),对照组(3只)不接种细胞。于接种第14天取部分小鼠处死,取脾脏,镜下观察其组织形态;接种第28天测算肿瘤体积。结果 LPS组、Poly(IC)组、Melan-A组成熟DC多于未处理组。接种DC+B16细胞的小鼠未见肿瘤生长;接种第28天,与接种B16细胞的小鼠比较,接种LPS、poly(IC)、Melan-A抗原肽激活的DC+B16细胞及接种B16细胞1周再接种DC+B16细胞的小鼠肿瘤体积减小(P均0.05)。与对照组比较,第14天接种DC+B16细胞小鼠脾脏滤泡结构无明显变化,而接种B16细胞及LPS、poly(IC)、Melan-A抗原肽激活的DC+B16细胞小鼠,脾脏滤泡结构增加。结论DC与B16共培养可抑制小鼠黑色素瘤的形成。
[Abstract]:Objective to investigate the effect of dendritic cell (DC) co-culture with melanoma cell line (B16) on the formation of mouse melanoma. Methods Murine bone marrow cells were extracted, differentiated and cultured to the 6th day. The cells were divided into 4 groups, lipopolysaccharide (LPS) group. In the Melan-A antigenic peptide group, the DC adjuvant LPS (final concentration 100 ng/m / L) was added to the polymyocytosis group. The final concentration was 20 ng/m / L, and the final concentration was 5 渭 mol / L (final concentration 5 渭 mol 路L ~ (-1)). No intervention was made in the untreated group. Flow cytometry was used to detect the mature state of DC. B16 cells were co-cultured with DC on the 6th day of culture for 2 days. Eight B16 cells were injected subcutaneously into C57BL / 6J mice. DC B16 cells, B16 cells were inoculated with DC B16 cells for 1 week, 5 DC B16 cells were inoculated, and 3 control cells were inoculated. Some of the mice were killed on the 14th day of inoculation, the spleen was taken and the histological morphology was observed under microscope. Results the number of mature DC in LPS group was more than that in untreated group. No tumor growth was found in mice inoculated with DC B16 cells, and 28 days after inoculation, compared with mice inoculated with B16 cells. The tumor volume of DC B16 cells inoculated with LPSN polyICN Melan-A antigen peptide and mice inoculated with DC B16 cells for 1 week decreased by 0.05. Compared with the control group, the splenic follicular structure of mice inoculated with DC B16 cells did not change significantly on the 14th day. The splenic follicular structure was increased in mice inoculated with B16 cells and activated by LPSN polyICN Melan-A antigenic peptide. Conclusion Co-culture of DC and B16 can inhibit the formation of melanoma in mice.
【作者单位】：天津医科大学基础医学院;天津医科大学药学院;
【基金】：国家自然科学基金资助项目(21373151)
【分类号】：R739.5

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