摘要
目的观察白桦酯酸作用前后对人CIK细胞增殖及对胃癌SGC-7901细胞杀伤活性的影响，探讨其发生的机制。方法分离健康者外周血单个核细胞（PBMC）在体外经多种细胞因子诱导为CIK细胞，收集培养第10天的CIK细胞，给予不同浓度白桦酯酸诱导48h，四甲基偶氮唑蓝（M1Tr）法检测人CIK细胞增殖率；流式细胞术（FCM）检测白桦酯酸作用前后CIK细胞穿孔素（PFP）、颗粒酶B（GrB）、CD107a的表达变化；乳酸脱氢酶（LDH）释放法测定白桦酯酸对CIK细胞杀伤胃癌细胞株SGC-7901的活性影响；Western blot检测药物诱导前后CIK细胞胞外信号调节激酶（ERKI／2）和接头蛋白76KD含有SH2结构域的白细胞特异性磷酸蛋白（SLP-76）、T细胞活化连接分子（LAT）的表达变化。结果白桦酯酸浓度在0．08～10μg／ml时能促进CIK细胞生长；经白桦酯酸诱导后的CIK细胞PFP、GrB、CD107a的表达显著高于对照组（P〈0．05），对胃癌SGC-7901细胞的杀伤活性亦显著高于对照组（P〈0．05）；经白桦酯酸作用的CIK细胞，其SLP-76、IJAT、ERKl／2的表达不同程度增加，显著高于对照组（P〈0．05）。结论白桦酯酸在一定浓度范围内能够促进CIK细胞增殖，并增强其对胃癌SGC-7901细胞的杀伤活性，其机制可能与活化SLP-76、LAT、ERK1／2，上调CIK细胞表面PFP、GrB、CD107a的表达有关。
Objective To observe the effect of betulinic acid（BetA） on the growth of human cyto- kine induced killer（CIK） ceils and the killing activity of CIK cells on the gastric cancer cells in vitro before and after induced by betulinic acid, explore its mechanism. Methods Peripheral blood mononuclear cell （PBMC） were separated form the healthy and were induced with various of cytokine to become CIK cells in vitro. CIK cells were collected on the tenth day and were induced with betulinic acid in different concentrations, followed by 48 h, the colorimetric methyl thiazolyl tetrazolium （MTT） method assay the proliferation rate of human CIK cells. Flow cytometry （ FCM ） was used to detect the expression changes of perforin, granzyme B and CD107a of human CIK cells before and after betulinic acid-induced. Lactate dehydrogenase （LDH） release assay was used to measure the influence on cytotoxic activity of CIK cells induced by betulin- ic acid against gastric cancer cell line SGC-7901 in vitro. Western blot assay was used to measure the extra- cellular signal-regulated kinasel/2 （ERK1/2）, and adapter proteins SH2-domain containing leukocyte pro- tein of 76KD（SLP-76） and linker for aetivative of T cells（LAT） expression changes of human CIK cells be-fore and after drug-induced. Results Betulinic acid can promote CIK cells growth when the concentration were in 0.08-10μg/ml, the expression of perforin, granzyme B and CD107a of CIK cells were siguificantly higher than control group（P〈0. 05 ） when the concentration of betulinic acid were in 0.3 μg/ml. In the meanwhile, the cytotoxic activity of CIK cells in vitro against gastric cancer cell line SGC-7901 were also remarkably higher than the control group（P〈0.05 ）. The expression of SLP-76, LAT and ERK1/2 were significantly increased to a certain extent than the control group（P〈0.05 ）, when CIK cells were treated with bet-ulinic acid. Conclusion These results suggest that betulinic acid can promote CIK ceils growth in some concentrations and increase the cytotoxic activity of CIK cells against gastric cancer cell line SGC-7901, its mechanism may related with two factors, on the one hand, enhancing the activity of SLP-76, LAT and ERK1/2, on the other hand, increasing the expression of perforin, granzyme B and CD107a on the surface of CIK ceils.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2012年第1期48-53,共6页
Chinese Journal of Microbiology and Immunology
基金
2009年度军区医学科技创新资助项目（09MA037）
关键词
白桦酯酸
胃癌细胞
CIK细胞
Betulinic acid
Gastric cancer cell line
CIK cells
