白藜芦醇对γδt细胞杀伤结肠癌sw1116细胞的影响及其机制研究-effect of resveratrol on γ δ t cells killing colon cancer sw 1116 cells and its mechanism.docxVIP

徐州医学院硕士学位论文白藜芦醇对丫6T细胞杀伤结肠癌SW-1116细胞 的影响及其机制研究中文摘要目的研究白藜芦醇(Resveratrol，Res)作用前后TST细胞对结肠癌SW-1116 细胞杀伤活性的变化并初步探讨其机制。方法异戊烯焦磷酸法体外扩增人外周血俩T细胞；不同浓度的白藜芦醇作 用于y6T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yfiT细胞及结肠癌SW-1116细胞生长的影响；流式细胞术(FCM)检测白藜芦 醇作用前后TST细胞穿孔素、颗粒酶B、CDl07a的表达；乳酸脱氢酶(LDH)释 放法检测白藜芦醇对Y6T细胞杀伤结肠癌SW-1116细胞的影响；Western．blot检测 白藜芦醇作用前后丫6T细胞细胞外信号调节激酶(ERKl／2)蛋白的活性。．结果浓度在0．39-3．1251xmol／L的白藜芦醇作用48h后，丫6T细胞的增殖率 较对照组显著提高(尸0．05)；浓度12．51001maol／L的白藜芦醇对结肠癌SW-1116 细胞生长有抑制作用，但无统计学差异；白藜芦醇作用后，TST细胞的穿孔素、颗 粒酶B、CDl07a的表达率显著高于对照组(P0．05)；浓度0．2～12．5I．tmol／L的白 藜芦醇作用48h后，对结肠癌细胞SW-1116的杀伤活性显著高于对照组(尸0．01)； 浓度O．1～lO岬ol／L白藜芦醇作用后，yST细胞的p-ERKl／2表达率较对照组显著增加(尸0．05)。结论适当浓度的白藜芦醇能够有效促进丫6T细胞的增殖，并增强其对结肠癌 SW-1116细胞的杀伤能力，其机制可能与白藜芦醇上调丫6T细胞穿孔素、颗粒酶B、 CDl07a的表达，活化细胞外信号调节激酶(p．ERKl／2)等有关。关键词白藜芦醇；结肠癌细胞：yST细胞2徐州医学院硕士学位论文Research of mechanism about enhancing the activity of yfT cells treated by resveratrol on colonic cancer cell lines SW．1116AbstractobjectiveTo explore the effect ofTfT cells on colonic cancer cell lines SW-1 1 16 treated by Resveratr01．MethodsAmplification丫6T cells of human Peripheral blood in vitro by usingisopentenylpyrophosphate．Different density Resveratrol induced 7fT cells and colonic cancelcell SW-1 1 16，methyl thiazolyl tetrazolium(M皿detectde the effect of growthand proliferation when different density Resveratrol action to Y6T cells and colonicCanCer cell lines SW-l l 1 6．Flow cytometer(FCM)detectde the expression of porforin，granzymeB and CD 1 07a on 7fT cells before it treated by Resveratrol and after that．7fT cells against colonic cancer cell lines SW-1 1 1 6 were detectde by lactate dehydrogenase(LDH)releasing assay before and after treated by Resveratr01．Western-blot analyzed the liveness of extracellular signal-regulated虹naSe 1／2 of TfT cells before it treated by Resveratrol and after that．ResultThe drug treatde on cells 48hours，when the density of Resveratrol in0．396--,3．125I_tmol／L yfTcells Was proliferation compared谢tll control group(尸0．05)． when the density of Resvemtrol in l 2．5-1 00I．