CAJ | 학술논문

目的探讨复方苦参对人γδT细胞杀伤胃癌细胞株SGC-7901的影响.方法用异戊烯焦磷酸法体外扩增人外周血γδT细胞,用不同浓度的复方苦参诱导γδT细胞和SGC-7901细胞株24h,用MTTDOI:10.3760/cma.j.issn.1673-4394.2009.04.004作者单位:221002,徐州医学院附属医院消化内科(韩露艳,费素娟);221004,解放军第97医院实验科(陈复兴,刘军权,陈桂林)通信作者:费素娟,E-mail:feisj1031@yahoo.com法检测复方苦参对这两种细胞生长抑制率的影响和LDH法检测γδT细胞的杀伤活性,用流式细胞术检测诱导前后的γδT细胞和SGC-7901的凋亡率.结果 γδT细胞培养10 d时从扩增前4.21%增加到70.35%,CD44达94.0%.不同浓度的复方苦参对SGC-7901细胞株的抑制率(22.3%)均明显高于γδT细胞(-22.4%),且γδT细胞的抑制率呈负的趋势,当复方苦参的浓度在1/50-1/400时γδT细胞负抑制率呈剂量依赖关系,且经复方苦参诱导24 h的γδT细胞杀伤活性(83.6%)明显高于先诱导SGC-7901组(71.2%),同时经复方苦参诱导24h的γδT细胞凋亡率(4.64%)明显低于SGC-7901(49.23%).结论复方苦参在临床常规使用浓度下,能够促进γδT细胞的增殖,同时能够抑制肿瘤细胞的生长,且能够增强γδT细胞的杀伤活性,这一结果将有助于肿瘤的过继免疫治疗及为复方苦参应用于肿瘤治疗提供了临床依据.
목적 탐토복방고삼대인γδT세포살상위암세포주SGC-7901적영향.방법 용이무희초린산법체외확증인외주혈γδT세포,용불동농도적복방고삼유도γδT세포화SGC-7901세포주24h,용MTTDOI:10.3760/cma.j.issn.1673-4394.2009.04.004작자단위:221002,서주의학원부속의원소화내과(한로염,비소연);221004,해방군제97의원실험과(진복흥,류군권,진계림)통신작자:비소연,E-mail:feisj1031@yahoo.com법검측복방고삼대저량충세포생장억제솔적영향화LDH법검측γδT세포적살상활성,용류식세포술검측유도전후적γδT세포화SGC-7901적조망솔.결과 γδT세포배양10 d시종확증전4.21%증가도70.35%,CD44체94.0%.불동농도적복방고삼대SGC-7901세포주적억제솔(22.3%)균명현고우γδT세포(-22.4%),차γδT세포적억제솔정부적추세,당복방고삼적농도재1/50-1/400시γδT세포부억제솔정제량의뢰관계,차경복방고삼유도24 h적γδT세포살상활성(83.6%)명현고우선유도SGC-7901조(71.2%),동시경복방고삼유도24h적γδT세포조망솔(4.64%)명현저우SGC-7901(49.23%).결론 복방고삼재림상상규사용농도하,능구촉진γδT세포적증식,동시능구억제종류세포적생장,차능구증강γδT세포적살상활성,저일결과장유조우종류적과계면역치료급위복방고삼응용우종류치료제공료림상의거.
Objective To explore the effect of Fufangkushen on gastric cancer cell killing by human γδT cells. Methods Isopentenyl pyrophosphate method was used to amplify human peripheral blood γδT cells in vitro. Fufangkushen at various concentrations was used to induce γδT cells and gastric cancer cell lines SGC-7901 for 24 hours, MTr assays was used to detect inhibitory effect of Fufangkushen on these cell lines, LDH assays was used to measure the cytotoxic activity of γδT cells, and flow cytometry was used to detect apoptosis of γδT cells and SGC-7901 before and after the treatment. Results Ten days after cultivation, proliferation ra-tio of γδT cells increased from 4.21% to 70.35% and CD44 was up to 94.0%. Inhibitory rate of Fufangkush-en on SGC-7901 at various concentrations was significantly higher than that on γδT cells (22.3% vs-22.4%, P<0.05). The negative inhibitory ratio on γδT cells showed a dose-dependent manner with Fufangkushen's concentrations ranging from 1/5 to 1/400. γδT cells cytotoxic activity to SGC-7901 induced by Fufangkushen for 24 h was higher than control group, (83.6% vs 71.2%, P<0.05). Apoptotic rate was significantly lower in γδT cells than in SGC-7901 (4.64% vs49.23%, P<0.05). Conclusion Fufangkushen, within routine concentration ranges, can promote γδT cells' proliferation, inhibit tumor cell growth and enhance γδT cells' cytotoxic activity. This may be beneficial to tumor adoptive immunotherapy and provide evidence for the appli-cation of Fufangkushen in the treatment of tumors.