HER2阳性乳腺癌细胞株上清液对树突状细胞表型及功能的影响

目的探讨人HER2阳性乳腺癌细胞对树突状细胞表型及功能的影响。方法分离培养外周血单个核细胞来源的树突状细胞。培养的第5天，实验组HER2’MCF-7细胞的培养上清液孵育24小时，以不加上清液的树突状细胞作为对照组，流式细胞术检测树突状细胞表面标志CDS0、CD86、CD40、CD83和HLA—DR的表达水平，ELISA法检测树突状细胞细胞因子IL-12、TNF—α、IL-6和IL-10的分泌水平。用改良的MTT比色法检测上清液处理的树突状细胞刺激同种异体T细胞增殖的能力和ELISA法检测树突状细胞刺激T细胞分泌细胞因子的能力。结果树突状细胞与HER2’MCF-7细胞的上清液共培养后，树突状细胞表面分子CD80、CD83、CD86及CD40均表达升高，同时伴随着IL-12、TNF-α、IL-6的分泌增加。与对照组相比较，上清液处理的树突状细胞对激活同种异体T细胞增殖和分泌细胞因子的能力明显增强（P〈0．01）。结论HER2’MCF-7细胞能改善树突状细胞的抗原递呈功能和刺激T细胞活化增殖的能力，提示HER2’MCF-7细胞对树突状细胞是免疫刺激作用，对于HER2’的乳腺癌可采用基于树突状细胞疫苗的免疫治疗。

The Effects of HER2+ MCF-7 Cell Supernatant on Phenotype and Function of Dendritic Cells

Objective To explore the effects of HER2＋MCF-7 cell supernatant on phenotype and function of dendrit- ic cells. Methods The peripheral blood mononuclear cells （PBMCs） were isolated and then cultured in RPMI-1640 with recombinant GM-CSF （10 ng/mL）, IL-4 （50 ng/mL） to obtain dendritic cells （DCs）. On day 5 of culture, DCs were treat- ed with HERUMCF-7 for 24 h. Supernatant- untreated DCs were used as control group. The expression of co-stimulato- ry molecules CDS0, CD86, CD40, CD83 and HLA-DR in DCs was evaluated by FACS, IL-12, TNF- α and IL-6 produc- tion by DCs was examined using ELISA. The proliferation and secretion of allogenetic T cells stimunlated by Supernatant- treated DCs were evaluated by MTT and ELISA. Results The levels of co-stimulatory molecules CDS0,CD86, CD40, CD83 and HLA-DR were elevated in DCs, accompanied by increased production of IL-12, TNF- c~, IL-6. Moreover, compared with the control group,HER2 ＋ MCF- 7- exposed DCs stimulated allogenetic T cell proliferation and T cell- derived cytokine production（P〈0.01）. Conclusions These results indicate that HERU MCF-7 can improve DCs＇ anti- gen presenting function and capability to activate T cells, suggesting that HER2＋ MCF-7 cells may have an immunostimu- latory effect on DCs, and DC based immunotherapy may be adopted in the treatment of breast cancer.