CAJ | 학술논문

目的探讨人结肠癌Lovo细胞总RNA抗原致敏的树突状细胞(DC)对细胞因子诱导的杀伤细胞(CIK)在体外特异性杀伤的影响.方法利用Ficoll密度梯度离心法提取脐血单核细胞,分别诱导CIK和DC细胞,并用流式细胞仪检测其免疫表型.采用Trizol提取结肠癌Lovo细胞总RNA作为肿瘤细胞抗原,转染脐血来源的DC.实验分为3组:转染Lovo DC共培养CIK组、未转染DC共培养CIK组和单纯CIK组.靶细胞为Lovo细胞,在效靶比为50∶1和20∶1的条件下以噻唑蓝法分别检测CIK的体外杀伤活性.结果在效靶比为20∶1时,负载Lovo RNA抗原的DC能诱导出CIK对Lovo细胞最强的细胞毒杀伤力为(76.49±4.21)%,DC+CIK组次之为(53.84±2.15)%,CIK组细胞毒性最低为(32.20±3.07)%,且两组间差异有统计学意义(P＜0.05).结论肿瘤细胞总RNA提取方法简单,易于临床实施,其作为抗原致敏DC能强化CIK的特异性杀伤,将有很好的临床应用前景.
목적 탐토인결장암Lovo세포총RNA항원치민적수돌상세포(DC)대세포인자유도적살상세포(CIK)재체외특이성살상적영향.방법 이용Ficoll밀도제도리심법제취제혈단핵세포,분별유도CIK화DC세포,병용류식세포의검측기면역표형.채용Trizol제취결장암Lovo세포총RNA작위종류세포항원,전염제혈래원적DC.실험분위3조:전염Lovo DC공배양CIK조、미전염DC공배양CIK조화단순CIK조.파세포위Lovo세포,재효파비위50∶1화20∶1적조건하이새서람법분별검측CIK적체외살상활성.결과 재효파비위20∶1시,부재Lovo RNA항원적DC능유도출CIK대Lovo세포최강적세포독살상력위(76.49±4.21)%,DC+CIK조차지위(53.84±2.15)%,CIK조세포독성최저위(32.20±3.07)%,차량조간차이유통계학의의(P＜0.05).결론 종류세포총RNA제취방법간단,역우림상실시,기작위항원치민DC능강화CIK적특이성살상,장유흔호적림상응용전경.
Objective To explore the effect of dendritic cells (DC) primed by total RNA extracted from human colon cancer Lovo cell on specific cytotoxicity of cytokine-induced killer cells (CIK) in vitro.Methods Cord blood mononuclear cells extracted by Ficoll density gradient centrifuge were induced into CIK and DC cells separately, and their Immunophenotype was detected by Flow cytometer. Trizol harvested total RNA from colon cancer cell Lovo and the RNAs were loaded to DCs obtained from cord blood as tumor anti gens. Effectors were grouped accordingly as CIK cells co-cultured with DCs transfected with Lovo RNA, CIK cells co- cultured with unloaded DCs and CIK cells. Targets was Lovo cells. In vitro cytotoxicity of CHK cells was extured with DCs loaded with Lovo RNA(76.49%±4.21%), DC + CIK group was lower(53.84% ± 2.15%),and CIK cells group possessed the lowest cytotoxicity(32.20% ± 3.07%), showing statistic significance( P ＜0.05). Conclusion Extraction of total RNA from tumor cells is simple and easy for clinical implementation.Total RNAs acted as antigen to pulse DCs can strengthen the specific cytotoxicity of CIK cells, which will have good prospects for clinical application.