舒林酸对人γδT细胞的作用研究

Research of Sulindac effect on human’s γδT cells

目的:探讨舒林酸在预防消化道肿瘤中对人γδT细胞的影响。方法:异戊烯焦磷酸法体外扩增人外周血γδT细胞。不同浓度的舒林酸诱导γδT细胞和胃腺癌细胞(SGC-7901),流式细胞术检测培养前后的γδT细胞表面标记及检测舒林酸诱导的γδT细胞和SGC-7901细胞凋亡百分率。乳酸脱氢酶法测定γδT细胞的杀伤活性。结果:γδT细胞培养10天时从扩增前4.21%增加到70.35%,CD44达94.0%。舒林酸在100%mol/L时对γδT细胞的生长抑制率达42.1%,明显高于对SGC-7901抑制率(7.4%)。γδT细胞和胃腺癌细胞SGC-7901细胞经不同浓度的舒林酸诱导24h后杀伤SGC-7901细胞作用在12.5μmol/L时最强。舒林酸浓度在100%mol/L时对γδT细胞作用24h的凋亡率(52.71%)显著高于SGC-7901细胞(7.88%)。结论:舒林酸在临床常规使用的药物浓度时可增强γδT细胞杀伤瘤细胞作用,超过这一浓度可明显抑制γδT细胞的增殖能力和杀伤活性,而对胃癌细胞株抑制作用不明显。这一结果为临床非甾体类药物预防消化道肿瘤的用量提供了实验依据。

Objective：To research the relationship of Sulindac and human＇s γδT cells on preventing digestive tract＇s tumor. Methods：Use the isopentenyl pyrophosphate method to amplify human peripheral blood γδT cells in vitro. After different concentration＇s Sulindac to induce γδT and SGC-7901 cells, flow cytometry was used to detect the cell surface marker of γδT cells before and after cultivation and the apoptosis percentage of γδT cells and SGC-7901 by Sulindac. Meanwhile, the killing and wounding activity of γδT cells was measured by lactate dehydrogenase method. Results：γδT cells were amplified from 4.21%to 70.35% in 10 days, CD44 up to 94.0%. when Sulindac＇s density is 100μmol/L, the growth inhibition ratio of γδT cells that up to 42.1% was notably higher than gastric carcinoma cell SGC-7901 ＇s growth inhibition ratio （7.4%）. After γδT cells and SGC-7901 induced by different concentration of Sulindac for 24 h, the cytotoxicity to SGC-7901 is the highest under 12.5 μmol/L Sulindac. The apoptosis percentage of γδT cells（52.71%） was significantly higher than SGC-7901＇s apoptosis percentage（7.88%） after induced by Sulindac（ 100μmol/L） for 24 h. Conclusion： Sulindac can increase the effect of γδT cells killing tumor cells in clinical routine drug concentration, while it can strikingly inhibit the reproductive and killing activity of γδT cells if its concentration higher than routine dose, but this inhibition effect is not obvious for gastric carcinoma cell. This result may offer an experimental base for applying clinical NSAIDs to prevent digestive tract＇s tumor.