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探讨水飞蓟宾对人γδT细胞杀伤胃癌细胞SGC-7901的影响及其作用机制。分离健康志愿者外周血单个核细胞,体外经多种细胞因子诱导培养为γδT细胞;收集培养、扩增7d后的γδT细胞,将其用不同浓度的水飞蓟宾诱导24h、48h及72h,CCK-8法检测γδT细胞增殖情况;流式细胞术(FCM)检测γδT穿孔素(perforin,PFP)、细胞颗粒酶B(Granzyme B,Gran B)及CDl07a的表达;western-blot法检测γδT细胞P-ERK1/2、Bcl-2、P-AKT、β-catenin的表达;乳酸脱氢酶(LDH)法检测γδT细胞对胃癌细胞SGC-7901的杀伤活性。结果发现浓度在1.6~100μg/ml的水飞蓟宾作用γδT细胞72h后,γδT细胞增殖率较对照组显著增加(P<0.05),且在25μg/ml时达到最高峰;将6.25~100μg/ml水飞蓟宾诱导γδT细胞72h后,γδT细胞的PFP、Gran B及CDl07a的表达以及P-ERK1/2、Bcl-2、P-AKT、β-catenin的表达与对照组比较均不同程度增加(P<0.05),且对胃癌细胞SGC-7901的杀伤活性显著增强(P<0.05)。以上实验结果提示一定浓度的水飞蓟宾对γδT细胞具有促进增殖作用,其机制可能与激活P-ERK1/2、Bcl-2、P-AKT及β-catenin信号通路有关。一定浓度的水飞蓟宾能够增加γδT细胞对胃癌细胞的杀伤活性,其作用机制可能与水飞蓟宾能够增加γδT细胞的穿孔素、Gran B及CDl07a的表达有关。
To investigate the effect of silibinin on human γδT cells killing SGC-7901 cells and its mechanism. The peripheral blood mononuclear cells from healthy volunteers were isolated and cultured in vitro to produce γδT cells induced by various cytokines. ΓδT cells cultured and expanded for 7 days were collected and induced with different concentrations of silybin for 24h, 48h and 72h, The proliferation of γδT cells was detected by CCK-8 assay. The expressions of γδT perforin (PFP), Granzyme B (Gran B) and CD107a were detected by flow cytometry (FCM) The expressions of P-ERK1 / 2, Bcl-2, P-AKT, and β-catenin in gastric cancer cell line SGC-7901 were detected by lactate dehydrogenase (LDH) assay. The results showed that the proliferation rate of γδT cells increased significantly (P <0.05) at the concentration of 1.6 ~ 100μg / ml after silybin was treated with γδT cells for 72h, reaching the peak at 25μg / ml; ml Silybin induced γδT cells for 72h, the expression of PFP, Gran B and CD107a and the expressions of P-ERK1 / 2, Bcl-2, P-AKT and β-catenin in γδT cells were increased to some extent compared with the control group (P <0.05), and the killing activity of SGC-7901 was significantly increased (P <0.05). The above results suggest that silybin at a certain concentration can promote the proliferation of γδT cells. The mechanism may be related to the activation of P-ERK1 / 2, Bcl-2, P-AKT and β-catenin signaling pathways. Silybin at a certain concentration could increase the cytotoxicity of γδT cells to gastric cancer cells, and its mechanism may be related to the increase of periostin, Gran B and CD107a expression of γδT cells by silybin.