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目的研究肝癌细胞(hepatocellular carcinoma cells,HCCs)微环境对树突状细胞(dendritic cells,DCs)的白介素-12(interleukin-12,IL-12)表达的影响,以进一步理解肿瘤的免疫逃逸机制。方法免疫磁珠法从人外周血分离CD14+单核细胞,加入rhGM-CSF和rhIL-4将单核细胞诱导分化为imDCs,利用rhTNF-α将imDCs诱导为mDCs,imDCs和mDCs与HCCs在Transwell中共培养48h,ELISA法分别检测不同培养条件下上清液中IL-10、IL-12、转化生长因子β1(trasnformed growth factor-β1,TGF-β1)和血管内皮生长因子(vascular endothelial growth factor,VEGF)的表达量。结果与HCCs共培养后,imDCs和mDCs的IL-12的表达量分别从(115.076±8.129)pg/ml和(258.346±3.609)pg/ml下调至(17.599±1.757)pg/ml和(6.787±1.123)pg/ml(P<0.05,P<0.01),而且培养上清中的TGF-β1浓度分别从(133.172±3.053)pg/ml和(174.446±3.972)pg/ml上调至(633.025±9.527)pg/ml和(486.806±10.515)pg/ml(P<0.05,P<0.01),VEGF的浓度分别从0pg/ml上调至(830.892±7.564)pg/ml和(858.169±5.419)pg/ml(P<0.01)。结论 HCCs可抑制DCs的IL-12的表达,这可能是HCCs损伤DCs免疫调节功能的方式之一。
Objective To investigate the effects of microenvironment of hepatocellular carcinoma cells (HCCs) on the expression of interleukin-12 (IL-12) in dendritic cells (DCs) to further understand the mechanism of tumor immune escape. Methods Immunomagnetic beads method was used to separate CD14 + monocytes from human peripheral blood. The monocytes were induced to differentiate into imDCs by adding rhGM-CSF and rhIL-4. The imDCs were induced to mDCs by rhTNF-α. The imDCs and mDCs were co- After cultured for 48 hours, the levels of IL-10, IL-12, TGF-β1 and VEGF in supernatants were detected by ELISA respectively ). Results After co-cultured with HCCs, the expression levels of IL-12 in imDCs and mDCs decreased from (115.076 ± 8.129) pg / ml and (258.346 ± 3.609) pg / ml to (17.599 ± 1.757) pg / ml and 1.123) pg / ml (P <0.05, P <0.01), and the TGF-β1 concentration in the culture supernatant was increased from (133.172 ± 3.053) pg / ml and (174.446 ± 3.972) pg / ml to (633.025 ± 9.527 ) were significantly increased from 0 pg / ml to (830.892 ± 7.564) pg / ml and (858.169 ± 5.419) pg / ml, respectively (P <0.01). Conclusion HCCs can inhibit the expression of IL-12 in DCs, which may be one of the ways that HCCs can impair the immunoregulatory function of DCs.