检验医学与临床Issue(15)：2161-2164,4.DOI:10.3969/j.issn.1672-9455.2015.15.009
负载抗原的DC与CIK共培养对多药耐药肝癌细胞HepG2／ADM的杀伤作用及其机制研究
Killing effects of co-culture of antigen-loaded DC and CIK cells on multidrug resistant liver cancer HepG2/ADM and its potential mechanism
摘要
Abstract
Objective To observe the killing effects of the antigen‐loaded dendritic cells(DC) and cytokine‐in‐duced killer cells(CIK ) on multidrug resistance liver cancer line HepG2/ADM with highly expressed permeability glycoprotein(P‐gp) and to explore its potential mechanisms .Methods DC cells and CIK cells were induced from the peripheral blood in healthy volunteers ′by the conventional methods ,and DC cells were impacted with HepG2/ADM cells after frozen‐thawed antigen ,which were co‐cultured with CIK cells for 24 ,48 ,72 ,96 h respectively .The DC cells without antigen‐loading and CIK cells were co‐cultured as control .The cell phenotype of DC and CIK was identified by the flow cytometry ,the cell proliferation vitality of HepG2/ADM was detected by CCK‐8 ;The expression of mdr‐1 mRNA was determined by realtime‐PCR ,the P‐gp protein level was detected by Western blot .Results Compared with non‐antigen‐loaded DC‐CIK group ,the expression of surface molecules in the antigen‐loaded DC‐CIK group was significantly increased(P< 0 .05) ,the vitality of HepG2 cells was inhibited more significantly (P < 0 .05) .The RT‐PCR and Western blot results showed that with the time extension ,the expression of mdr‐1 mRNA and P‐gp protein both were significantly decreased in the above two groups respectively(P< 0 .05) ;Furthermore ,the inhibitory effects was more significant in the antigen‐loaded group(P< 0 .05) .Conclusion The co‐culture of DC and CIK after antigen impact could improve its killing activity to multidrug‐resistant HepG2/ADM cell line ,its potential mechanism may be via inhibiting the MDR closely related mdr‐1 gene expression and its encoded P‐gp protein level .关键词
细胞因子诱导的杀伤细胞/树突状细胞/多药耐药/P-糖蛋白/HepG2/ADM 细胞Key words
cytokine-induced killer cells/dendritic cells/multidrug-resistance/P-gp/HepG2/ADM cells引用本文复制引用
徐巧元,杨志祥,罗阔..负载抗原的DC与CIK共培养对多药耐药肝癌细胞HepG2／ADM的杀伤作用及其机制研究[J].检验医学与临床,2015,(15):2161-2164,4.基金项目
重庆市渝中区科技计划项目（20120225）。 （）
