具有肿瘤杀伤活性粒细胞的筛选及其体外抑瘤效应观察  

Screening of granulocytes with cancer killing activity and their in vitro anti-tumor effects

作　　者：郭慧军[1] 赵树铭[1] 

机构地区：[1]第三军医大学西南医院输血科,重庆400038

出　　处：《第二军医大学学报》2012年第5期474-478,共5页Academic Journal of Second Military Medical University

摘　　要：目的探索具有肿瘤杀伤活性(CKA)粒细胞的筛选方法,并对其体外抑瘤效应进行初步研究。方法 (1)收集健康志愿者外周抗凝全血21份,分离粒细胞,与人肺癌A549细胞共培养,通过检测肿瘤细胞增殖活性筛选出CKA粒细胞。(2)将筛选得到的CKA粒细胞作用于不同种类的肿瘤细胞,采用CCK-8法检测肿瘤细胞活性,双染法检测肿瘤细胞凋亡发生情况。(3)分别采用直接接触、Transwell共培养、条件培养液等共培养细胞方式,观察不同处理方式对CKA粒细胞抑制肿瘤细胞活性的影响。结果 (1)有6株粒细胞克隆对A549细胞活性具有明显的抑制作用,将其中作用最显著的克隆3标记为CKA粒细胞。(2)CKA粒细胞不仅对人肺腺癌细胞A549、人肝癌细胞HepG2和人宫颈癌细胞HeLa生长活性有明显抑制作用,且能促进这3种细胞的凋亡。(3)3种处理方式的抑制效率比较:直接接触>Transwell>条件培养。结论 CKA粒细胞对某些类型的肿瘤细胞活性具有明显的抑制作用;CKA粒细胞对肿瘤的杀伤作用可能与其与肿瘤的接触方式有关。Objective To explore the screening method for granulocytes with cancer killing activity（CKA） and to observe their in vitro anti-tumor effects.Methods（1） Anti-coagulated peripheral whole blood samples were collected from 21 healthy volunteers and were co-cultured with human lung cancer cell line A549.The proliferation activity of cancer cells was observed to screen granulocytes with CKA.（2） The screened granulocytes with CKA was used to treat various tumor cells,CCK-8 method was used to examine the vitality of tumor cells,and the cell apoptosis was detected by Annexin Ⅴ-FITC Apoptosis Detection Kit Ⅱ.（3） The cancer cells were co-cultured using direct contact,Transwell and conditioned medium,and the effects of the three methods on the anti-tumor effect of CKA granulocytes were observed.Results（1） Six granulocyte clones showed noticeable inhibitory effect against A549 cells,and clone 3 had the most potent effect and was marked as CKA granulocyte.（2） The screened CKA granulocytes not only had noticeable inhibitory effects against A549,HepG2 and HeLa cells,but also promoted their apoptosis.（3）The order of the inhibitory efficiencies for the three culture methods from strong to weak was： direct contact Transwell conditioned medium.Conclusion CKA granulocytes have noticeable inhibitory effect against some cancer cells,which might be associated with their contact mode with the cancer cells.

关 键 词：肿瘤杀伤活性 粒细胞 筛选 肿瘤治疗 

分 类 号：R73-36[医药卫生—肿瘤]