吉西他滨联合树突状细胞对HepG2 细胞增殖的抑制作用

J4 ›› 2011, Vol. 37 ›› Issue (3): 460-464.

吉西他滨联合树突状细胞对HepG2 细胞增殖的抑制作用

付京| 陈域| 雷海路| 雷团结| 张鹏

中国人民解放军第451医院肝胆普外科|陕西西安 710054

收稿日期:2011-04-07 出版日期:2011-05-28 发布日期:2011-05-28
作者简介:付京（1960-）|男|北京市人|副主任医师|主要从事普外科疾病的基础及临床研究。
基金资助:
陕西省科技厅科技攻关项目资助课题（2008k-02）

Inhibitory effect of gemcitabine combined with dendritic cells on proliferation of HepG2 cells

FUJing,CHEN Yu,LEI Hai-lu,LEI Tuan-jie,ZHANG Peng

Department of Hepatobiliary General Surgery,No.451 Hospital of PLA,Xi’an 710054,China

Received:2011-04-07 Online:2011-05-28 Published:2011-05-28

摘要/Abstract

摘要：

目的：观察肿瘤细胞裂解物致敏的树突状细胞（DC）联合吉西他滨（GEM）体外抑制肝癌HepG2 细胞系增殖的作用，为临床探索有效的肝癌综合治疗方案提供实验依据。方法：采用反复冻融法裂解HepG2 细胞，提取肿瘤细胞抗原后致敏DC，并以DC刺激自身T 细胞的增殖和活化。按不同处理因素分组：①空白对照组，只含空白培养液；②阴性对照组，只含HepG2 细胞；③单纯T 细胞组，将未经致敏DC 刺激的T细胞与靶细胞按20∶1 的比例接种；④单纯活化T 细胞组，将经致敏DC 刺激后的活化T 细胞与靶细胞按20∶1 的比例接种；⑤单纯GEM 处理组，靶细胞接种后在培养基中分别添加终浓度为50、100、150、200 μg/L^-1 的GEM；⑥T细胞+GEM 处理组，在单纯T 细胞组的培养基中分别添加上述4个浓度梯度的GEM；⑦活化T 细胞+GEM 处理组，在活化T 细胞组的培养基中分别添加上述4个浓度梯度的GEM。MTT 法检测细胞杀伤率，流式细胞仪检测细胞凋亡率，观察应用T 细胞、不同浓度的GEM 以及两者联合应用对HepG2 细胞的体外杀伤效应。结果：空白对照组刺激指数（SI）为1.34，阴性对照组SI 为3.48，两者比较差异有统计学意义（P<0.01）。与单纯T细胞组［（4.81±2.54）%］比较，单纯活化T 细胞组细胞杀伤率［（21.68±4.39）%］明显升高（P<0.01）；与活化T细胞+50、100 和150 μg/L^-1 GEM组比较，活化T细胞+200 μg/L^-1 GEM组细胞杀伤率明显升高（P<0.05）。单纯T 细胞组细胞凋亡率为（5.45±0.94）%，与阴性对照组［（4.47±1.98）%］比较差异无统计学意义（P>0.05）；活化T 细胞组细胞凋亡率为（14.32±1.16）%，与前两组比较差异有统计学意义（P<0.01）。活化T细胞+100 μg/L^-1 GEM组细胞凋亡率为（24.52±0.85）%，与其他各组比较差异均有统计学意义（P<0.05）。结论：负载肿瘤细胞抗原的DC 可诱导出抗肿瘤的细胞毒性T细胞，活化的T细胞联合GEM 能显著提高其杀伤肿瘤细胞的作用。

关键词: 原发性肝癌；树突状细胞；吉西他滨

Abstract:

Abstract:Objective To observe the inhibitory effect of gemcitabine (GEM) combined with the dendritic cells (DC) sensitized byHepG2 cells’ whole lysates on proliferation of HepG2 cells and provide experimental basis for its clinical application.Methods Whole cell lysates of HepG2 cells were extracted with freeze thawing method,then the monocyte-derived DC were plusedinto this cellular antigen. The HepG2 cells were divided into negative group(only HepG2 cells);unactivated T lymphocytes(UTL) group;activated T lymphocytes(ATLgroup ;50, 100,150,and 200 μg/L^-1 GEM groups;UTL+ 50,100,150,and200 μg/L^-1 GEM groups;ATL+50,100,150,and 200 μg/L^-1 GEM groups.The capability of antigen plused DC to promote T lymphocyte proliferation and the abilities of different concentrations of GEM combined with sensitized T cells to kill HepG2 cells were detected by MTT methods,respectively. The apoptotic rates of HepG2cells in various groups were detected by flow cytometry. Results There was significant difference of the stimulate index(SI) between control and negative control groups（P<0.01）.The kill rate in ATL group(21.68%±4.39%) was higher than thatin UTL group （4.81%2.54%）,there was significant difference（P<0.01）.Compared with ATL+50,100,150 μg/L^-1 GEgroups,the kill rate in ATL+ 200 μg?L-1 GEM group was significantly increased（P<0.05）.Compared with negative control group（4.47%±1.98%）,the apoptotic rate in UTL group （5.45%±0.94%)had no significant differenc（P>0.05） ;the apoptotic rate in ATL group （14.32%±1.16%）had significant differences compared with the former two groups（P<0.01）. The apoptotic rate in ATL+ 100 μg/L^-1 GEM group was 24.52%±0.85%,there were significant differences compared with the other groups（P<0.05）. Conclusion The DC sensitized by HepG2 cells’ whole lysates could induce anti-cancer cytotoxicity T cells and the activated T cells combined with GEM can significantly increase their effects to kill tumor cells .

Key words: primary liver carcinoma;dendritic cell;gemcitabine

中图分类号:

R735.7

付京|陈域|雷海路|雷团结|张鹏. 吉西他滨联合树突状细胞对HepG2 细胞增殖的抑制作用[J]. J4, 2011, 37(3): 460-464.

FUJing,CHEN Yu,LEI Hai-lu,LEI Tuan-jie,ZHANG Peng . Inhibitory effect of gemcitabine combined with dendritic cells on proliferation of HepG2 cells[J]. J4, 2011, 37(3): 460-464.

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