大鼠胰腺癌微环境诱导不成熟树突状细胞功能变化的实验研究

发布时间：2018-05-26 05:07

本文选题：树突状细胞 + 胰腺癌微环境　；参考：《山西医科大学》2011年硕士论文

【摘要】：目的: 初步研究胰腺癌微环境对树突状细胞(Dendritic cell，Dc)成熟的影响及功能变化并初步探讨胰腺肿瘤细胞免疫逃逸的发生机制。方法：培养树突状细胞，加入粒细胞巨噬细胞集落刺激因子(GM-CSF)40ng/ml、白细胞介素4(IL-4)40ng/ml，培养到第６天时得到大量的未成熟树突状细胞(immaturedendriticcells,imDC)，，加入大鼠胰腺癌细胞（AR42J cell）培养上清液诱导共培养3天，流式细胞术检测DC的表面分子CD86、CD80的表达（n=6），观察能否延缓或阻断imDC的成熟及其在脂多糖(lipopolysaccharide,LPS)刺激后这种作用能否被逆转。并观察AR42J细胞培养上清诱导的DC对同种异体混合淋巴细胞增殖情况。结果：加入AR42J细胞培养上清液诱导的DC与正常培养的DC比较，CD80+ CD86+阳性率分别为（7.15±0.71）%和（70.88±3.60）%，AR42J细胞培养上清诱导的DC经LPS刺激后细胞的表面分子CD80+CD86+阳性率表达仍然较低（7.43±1.05）%，表明AR42J细胞培养上清液对imDC的成熟有阻断作用。AR42J细胞培养上清液诱导的imDC组刺激同种异体混合淋巴细胞增殖的强度显著低于正常培养的imDC组刺激同种异体混合淋巴细胞增殖的强度。结论: 体外AR42J细胞培养上清液可以诱导imDC处于不成熟状态，这种不成熟状态不容易被逆转,并且这种被诱导的imDC与正常培养的imDC比较刺激同种异体混合淋巴细胞增殖的能力显著降低。
[Abstract]:Objective: To study the effect of pancreatic cancer microenvironment on dendritic cell (DC) maturation and its functional changes, and to explore the mechanism of pancreatic tumor cell immune escape. Methods: Dendritic cells were cultured, then 40 ng / ml of granulocyte macrophage colony stimulating factor (GM-CSFN) and 40 ng / ml of interleukin-4 (IL-4) were added into the culture medium. On the 6th day, a large number of immature dendritic cells were obtained and cultured in the supernatant of rat pancreatic cancer cell line AR42J for 3 days. Flow cytometry was used to detect the expression of CD86 + CD80 on the surface of DC and to observe whether it could delay or block the maturation of imDC and whether it could be reversed after lipopolysaccharide (LPS) was stimulated by lipopolysaccharide. The proliferation of allogeneic mixed lymphocytes induced by supernatants of AR42J cells was observed. Results: The positive rates of CD80 CD86 were 7.15 卤0.71% and 70.88 卤3.60% in DC induced by AR42J cell culture supernatant and 70.88 卤3.60% in normal DC, respectively. The CD80 CD86 positive rate of DC induced by LPS was still lower than that of DC cultured in normal culture. The supernatant of AR42J cell culture can block the maturation of imDC. The intensity of stimulating the proliferation of allogeneic mixed lymphocytes in imDC group induced by supernatant of AR42J cell culture is significantly lower than that in imDC group of normal culture. The intensity of cell proliferation. Conclusion: The supernatant of AR42J cell culture in vitro could induce imDC to be in immature state, which could not be easily reversed, and the ability of inducing imDC to stimulate the proliferation of allogeneic mixed lymphocytes was significantly lower than that of normal cultured imDC.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R392

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