PBMC与HepG2细胞体外共培养时的相互作用研究  

The Interaction between PBMC and HepG2 during In Vitro Coculture

作　　者：邵红伟[1] 苏畅[1] 李侃毅 沈晗[1] 张文峰[1] 吴凤麟[1] 王辉[1] 薄华本[1] 黄树林[1] 

机构地区：[1]广东药学院生命科学与生物制药学院/广东省生物技术候选药物研究重点实验室,广东广州510006

出　　处：《药物生物技术》2015年第4期293-297,共5页Pharmaceutical Biotechnology

基　　金：国家自然科学基金基金(No.31100664;31300737;81303292);广东省自然科学基金(No.10151022401000024;2014A030313586);广东药学院师资队伍建设专项经费

摘　　要：淋巴细胞与肿瘤细胞之间存在着复杂的相互调控和抑制作用,作为细胞分泌的一种囊泡,外泌体(Exosome)能够发挥调控细胞生长和活性的作用。该研究将分离自健康人的外周血单核细胞(Peripheral blood mononuclear cells,PBMC)与肿瘤细胞Hep G2共培养,以检测两者的相互作用以及细胞因子和外泌体的分泌情况。结果表明在一定的范围内肿瘤细胞越多,刺激淋巴细胞分泌IFN-γ的水平越高;流式和激光共聚焦检测显示两者可直接吸附或者通过产生外泌体附着在对方表面;利用两者含有外泌体的培养上清进行交互处理,流式检测表明超过30%的PBMC被肿瘤细胞外泌体所吸附,活化的PBMC更易被肿瘤细胞外泌体吸附,而肿瘤细胞则较少被PBMC外泌体吸附;并且活化的淋巴细胞经肿瘤细胞外泌体处理后,活化程度显著降低,而肿瘤细胞受到淋巴细胞外泌体的影响则较小。这些结果表明淋巴细胞主要通过直接接触发挥作用,而肿瘤细胞则更多地通过外泌体发挥作用,这为更进一步的了解淋巴细胞和肿瘤细胞之间的相互作用机制,制定新的抗肿瘤策略奠定了基础。There is a complex mutual regulation and inhibition of lymphocytes and tumor cells in tumor microenvironment. Exosome,nanometer-sized vesicles released from a wide variety of cells,can regulate the development and activity of cells and has attracted more and more attention in the immune regulation of tumor cells. In the present study,peripheral blood mononuclear cells（ PBMC）isolated from healthy individual were co-cultured in vitro with tumor cells Hep G2 to investigate the interaction of them and the production of cytokines and exosomes. It was showed that in a certain range of effect to target ratio（ E： T） the more tumor cells there were,the more IFN-γ released by PBMC. With flow cytometry（ FCM） analysis and confocal laser scanning microscopy（ CLSM） direct observation,PBMC and Hep G2 cells were found to adsorb each other directly or release exosomes to target each other. Treated with exosome-containing culture supernatant of Hep G2 cells,more than 30% of PBMC were labeled with tumor cell-derived exosomes. The percentage is higher in activated PBMC. However,treated with exosome-containing culture supernatant of PBMC,very few tumor cells were labeled with the PBMC-derived exosomes. Furthermore,activated lymphocytes treated with tumor cell-derived exosomes showed significantly reduced activity,while tumor cells treated with lymphocytes-derived exosomes showed no significant change in the secretion of TGF-β. These results indicated that lymphocytes exert effect on tumor cells mainly through cell-cell contact,while tumor cells excert effect on lymphocytes more likely through releasing exosomes,which laid a foundation for further exploring the interaction mechanism between lymphocytes and tumor cells and developing novel anti-tumor strategies.

关 键 词：外周血单核细胞 肿瘤细胞 相互作用 外泌体 吸附 细胞因子 

分 类 号：Q256[生物学—细胞生物学]