| 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CCK8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。 |
| 关键词: 肿瘤杀伤活性 粒细胞 筛选 肿瘤治疗 |
| DOI：10.3724/SP.J.1008.2012.00474 |
| 投稿时间：2012-04-24修订日期：2012-05-04 |
| 基金项目: |
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| Screening of granulocytes with cancer killing activity and their in vitro anti tumor effects |
| GUO Hui-jun,ZHAO Shu-ming* |
(Department of Blood Transfusion, Southwest Hospital, Third Military Medical University, Chongqing 400038, China
*Corresponding author.) |
| Abstract: |
| ObjectiveTo explore the screening method for granulocytes with cancer killing activity (CKA) and to observe their in vitro antitumor effects. Methods(1) Anticoagulated peripheral whole blood samples were collected from 21 healthy volunteers and were cocultured 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, CCK8 method was used to examine the vitality of tumor cells, and the cell apoptosis was detected by Annexin ⅤFITC Apoptosis Detection Kit Ⅱ. (3) The cancer cells were cocultured using direct contact, Transwell and conditioned medium, and the effects of the three methods on the antitumor 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. |
| Key words: cancer killing activity granulocytes screening oncotherapy |
