Objective： The aim of our study is to observe the impact of cholangiocarcinoma-derived exosomes on the antitumor activities of cytokine-induced killer （CIK） cells and then demonstrate the appropriate mechanism. Met...Objective： The aim of our study is to observe the impact of cholangiocarcinoma-derived exosomes on the antitumor activities of cytokine-induced killer （CIK） cells and then demonstrate the appropriate mechanism. Methods： Tumor-derived exosomes （TEXs）, which are derived from RBE cells （human cholangiocarcinoma line）, were collected by ultracentrifugation. CIK cells induced from peripheral blood were stimulated by TEXs. Fluorescence-activated cell sorting （FACS） was performed to determine the phenotypes of TEX-CIK and N-CIK （normal CIK） cells. The concen- trations of tumor necrosis factor-a （TNF-a） and perforin in the culture medium supematant were examined by using an enzyme-linked immunosorbent assay （ELISA） kit. A CCK-8 kit was used to evaluate the cytotoxic activity of the CIK cells to the RBE cell line. Results： The concentrations of TNF-a and perforin of the group TEX-CIK were 138.61 pg/ml and 2.41 ng/ml, respectively, lower than those of the group N-CIK 194.08 pg/ml （P〈0.01） and 3.39 ng/ml （P〈0.05）. The killing rate of the group TEX-CIK was 33.35%, lower than that of the group N-CIK （47,35% （P〈0.01））. The population of CD3＋, CD8＋, NK （CD56＋）, and CD3＋CD56＋ cells decreased in the TEX-CIK group （63.2±6.8）%, （2.5±1.0）%, （0.53±0.49）%, （0.45±0.42）%） compared with the N-CIK group （（90.3±7.3）%, （65.7±3.3）%, （4.2±1.2）%, （15.2±2.7）%）, P〈0.01. Conclusions： Our results suggest that RBE cells-derived exosomes inhibit the antitumor activity of CIK cells by down-regulating the population of CD3＋, CD8＋, NK （CD56＋）, and CD3＋CD56＋ cells and the secretion of TNF-a and perforin. TEX may play an important role in cholangiocarcinoma immune escape.展开更多
基金Project supported by the National Natural Science Foundation of China(No.81272671)the Foundation of Health and Family Planning Commission of Zhejiang Province(No.2015KYB218)China
文摘Objective： The aim of our study is to observe the impact of cholangiocarcinoma-derived exosomes on the antitumor activities of cytokine-induced killer （CIK） cells and then demonstrate the appropriate mechanism. Methods： Tumor-derived exosomes （TEXs）, which are derived from RBE cells （human cholangiocarcinoma line）, were collected by ultracentrifugation. CIK cells induced from peripheral blood were stimulated by TEXs. Fluorescence-activated cell sorting （FACS） was performed to determine the phenotypes of TEX-CIK and N-CIK （normal CIK） cells. The concen- trations of tumor necrosis factor-a （TNF-a） and perforin in the culture medium supematant were examined by using an enzyme-linked immunosorbent assay （ELISA） kit. A CCK-8 kit was used to evaluate the cytotoxic activity of the CIK cells to the RBE cell line. Results： The concentrations of TNF-a and perforin of the group TEX-CIK were 138.61 pg/ml and 2.41 ng/ml, respectively, lower than those of the group N-CIK 194.08 pg/ml （P〈0.01） and 3.39 ng/ml （P〈0.05）. The killing rate of the group TEX-CIK was 33.35%, lower than that of the group N-CIK （47,35% （P〈0.01））. The population of CD3＋, CD8＋, NK （CD56＋）, and CD3＋CD56＋ cells decreased in the TEX-CIK group （63.2±6.8）%, （2.5±1.0）%, （0.53±0.49）%, （0.45±0.42）%） compared with the N-CIK group （（90.3±7.3）%, （65.7±3.3）%, （4.2±1.2）%, （15.2±2.7）%）, P〈0.01. Conclusions： Our results suggest that RBE cells-derived exosomes inhibit the antitumor activity of CIK cells by down-regulating the population of CD3＋, CD8＋, NK （CD56＋）, and CD3＋CD56＋ cells and the secretion of TNF-a and perforin. TEX may play an important role in cholangiocarcinoma immune escape.
