乌司他丁联合多西他赛对乳腺癌免疫微环境的影响

Effect of ulinastatin and docetaxel on immune microenvironment of breast cancer

摘要:

目的：探讨乌司他丁（ulinastatin，UTI）联合多西他赛（docetaxel，DCT）对乳腺癌微环境中免疫细胞，免疫因子和基质的作用，以及对肿瘤生长的影响。方法：进行4T1乳腺癌细胞株培养，购买BALB/c雌性小鼠，建立BALB/c小鼠荷4T1细胞模型，将32只小鼠荷瘤随机分为4组，对照组，UTI组，DCT组，UTI+DCT组。测量各组肿瘤体积，计算肿瘤抑制率；行流式细胞染色，检测CD4+CD25+Foxp3+Treg的含量；ELISA检测白介素（interleukin，IL）2、IL-6和IL-10的表达；Western blot检测基质金属蛋白酶9（matrix metalloproteinases 9，MMP-9）以及金属蛋白酶组织抑制因子1（tissue inhibitor of metalloproteinase 1，TIMP1）。结果：与对照组相比，UTI和DCT均能抑制肿瘤生长，UTI联合DCT作用最为明显（P=0.000），UTI、DCT单独和联合用药后，CD4+CD25+Foxp3+Treg/CD4+T细胞的比值较对照组降低（P=0.000），IL-6、IL-10和MMP-9的表达较对照组明显下降（P=0.000），同时促进IL-2及TIMP1的表达（P=0.000）。结论：UTI联合DCT可抑制免疫正常的小鼠乳腺癌的生长，其机制可能与调节肿瘤微环境中CD4+CD25+Foxp3+Treg、IL-2、IL-6、IL-10、MMP-9 以及TIMP1有关。

Abstract:

Objective：To investigate the effect of ulinastatin（UTI） and docetaxel（DCT） on immunocyte，immune factors and matrixes in breast cancer immune microenvironment and the tumor growth. Methods：4T1 breast cancer cells were cultured and the BALB/c mice were purchased to establish BALB/c mice model bearing 4T1 cells. Thirty-two mice were randomly divided into four groups：control group，UTI group，DCT group and UTI + DCT group. The tumor size was measured and the inhibition rate was calculated. Line flow cytometric dyeing was used to detect CD4+CD25+Foxp3+Treg content. The expression of interleukin-2（IL-2），IL-6 and IL-10 was detected by ELISA. The expression of matrix metalloproteinases 9（MMP-9） and tissue inhibitor of metalloproteinase 1（TIMP1） was detected by Western blot. Results：Both UTI and DCT can inhibit tumor growth and inhibitory effect was the most significant when UTI combined with DCT（P=0.000）. The rate of CD4+CD25+Foxp3+Treg/CD4+T cells was decreased in UTI group，DCT group and UTI + DCT group than in control group（P=0.000）. The expression of IL-6，IL-10 and MMP-9 was significantly decreased and the expres－sion of IL-2 and TIMP1 was increased in UTI group，DCT group and UTI + DCT group than in control group（P=0.000，P=0.000）. Conclusion：UTI combined with DCT can suppress the tumor growth in mice with normal immune function. The mechanism may be related with regulating CD4+CD25+Foxp3+Treg，IL-2，IL-6，IL-10，MMP-9 and TIMP1 in the tumor microenvironment.