白藜芦醇对γδT细胞杀伤结肠癌SW-1116细胞的影响及机制研究

Research of mechanism about enhancing the activity of vST cells treated by resveratrol on colonic cancer cell lines SW-1116

目的观察白藜芦醇作用前后γδT细胞对结肠癌SW-1116细胞杀伤活性的变化，并探讨其发生的机制。方法异戊烯焦磷酸法体外扩增人外周血γδT细胞，不同浓度的白藜芦醇作用于γδT细胞和结肠癌SW-1116细胞，四甲基偶氮唑蓝（MTT）法检测白藜芦醇对γδT细胞及结肠癌细胞的生长的影响；流式细胞术（FCM）检测白藜芦醇作用前后γδT细胞穿孔素、颗粒酶B、CD107a的表达；乳酸脱氢酶（LDH）释放法检测白藜芦醇对γδT细胞杀伤结肠癌SW-1116细胞活性的影响。Westernblot检测药物作用前后γδT细胞细胞外信号调节激酶（ERK1／2）蛋白的活性的变化。结果白藜芦醇在0．39～3．125tzmol／L时对75T细胞的生长具有促进作用，对结肠癌SW—1116细胞作用不明显；经白藜芦醇诱导后γδT细胞的穿孔素、颗粒酶B、CD107a的表达显著高于对照组（P〈0．05）；对结肠癌SW-1116细胞的杀伤活性也显著高于未诱导组（P〈0．05）；经浓度为0．1—10μmol／L白藜芦醇作用的18T细胞的p-ERK1／2表达较对照组增加（P〈0．05）。结论白藜芦醇能够促进18T细胞的增殖，并增强其对结肠癌SW-1116细胞的杀伤能力，其机制可能与上调γδT细胞表面的穿孔素、颗粒酶B、CD107a的表达及活化细胞外信号调节激酶等有关。

Objective To explore the effect of γδT cells on colonic cancer cell lines SW-1116 treated by resveratrol. Methods Amplification γδT cells of human peripheral blood in vitro by using isopentenylpyro- phosphate. Different density resveratrol induced γδT cells and colonic cancer cell SW-1116, methyl thiazolyl tetrazolium（MTF） detectde the effect of growth and proliferation when expression veratrol action to γδT cells and colonic cancer cell lines SW-1116. Flow eytometer （ FCM ） detectde the expression of porforin, granzymeB and CD107a on γδT cells before it treated by resveratrol and after that. γδT cells against colonic cancer cell lines SW-1116 were detectde by lactate dehydrogenase（LDH） releasing assay before and after treated by resveratrol. Western blot analyzed the liveness of extraeellular signal-regulated kinasel/2 of γδT cells before it treated by resveratrol and after that. Results γδTcells would be proliferation when the density of resveratrol in 0.39- 3. 125 μmol/L, while the effect on colonic cancer cell lines SW-1116 was not significance. There was significant difference of the expression of porforin, ranzymeB and CD107a on γδT cells before it treated by resveratrol and after that （P〈0.05）. γδT cells against colonic cancer cell lines SW-1116 treated by resveratrol was enhanced compared with control group（P〈0.05） . The p-ERK 1/2expression of γδT ceils enhanced when the density of resveratrol treated to γδT cells in 0. 1-10 p.mol/L （P〈 0.05 ）. Conclusion Resveratrol could promote γδT cells, proliferation and strengthen the cytotoxicity of γδT cells against colonic cancer cell SW-1116, the mecha- nism might concerned with activating p-ERK1/2 and enhancing the expression of porforin, granzymeB and CD107a on γδT cells.