三裂鼠尾草素对人γδT细胞杀伤结肠癌SW-620细胞的影响

Effects of salvigenin on human γδT cell killing of colon cancer cell lines SW-620

目的探讨三裂鼠尾草素对人γδT细胞杀伤结肠癌SW-620细胞的影响。方法分离健康人外周血单个核细胞,异戊烯焦磷酸法体外扩增γδT细胞,不同浓度三裂鼠尾草素诱导人γδT细胞72 h,CCK8法检测γδT细胞增殖率,乳酸脱氢酶释放法检测诱导后γδT细胞对SW-620细胞的杀伤活性,流式细胞术检测诱导后γδT细胞穿孔素（PFP）、颗粒酶B（Gra B）、CD107a的表达,Western blot检测诱导后γδT细胞p-ERK1/2的表达情况。结果经培养10 d,γδT细胞比例由4.32%增加至83.63%。三裂鼠尾草素浓度为0.047 7-195 pmol/L时γδT细胞增殖率显著升高（P〈0.05）,浓度为0.191-195 pmol/L时γδT细胞对SW-620细胞的杀伤活性显著增加（P〈0.05）。经三裂鼠尾草素诱导72 h,γδT细胞PFP、Gra B、CD107a的表达显著升高（P〈0.05）。三裂鼠尾草素浓度为0.763-195 pmol/L时γδT细胞p-ERK1/2的表达显著增加（P〈0.05）。结论在一定浓度范围内三裂鼠尾草素能够促进人γδT细胞增殖,增强其对结肠癌细胞SW-620的杀伤活性,其机制可能与PFP、Gra B的表达上调和p-ERK1/2信号通路活化有关。

Objective To investigate the effects of salvigenin on human γδT cell killing of colon cancer cell lines SW -620. Methods Peripheral blood mononuclear cells were separated from healthy subjects and isopentenyl pyrophosphate method was used to amplify γδT cells in vitro. After γδT cells were cultured with different concentrations of salvigenin for 72 h, CCK8 assay was used to assess the proliferation of γδT cells. Lactate dehydrogenase release assay was applied to measure the cytotoxic activity of γδT cells to colon cancer cells in vitro. Flow cytometry was performed to detect the expression of granzyme B （GraB）, perforin （PFP） and CD107a of γδT cells after treatment. Western blot assay was used to test p - ERK1/2 expression in the treated γδTT cells. Results After ten days of culture, the proportion of γδT cells increased from 4. 32% to 83.63%. When salvigenin concentration was between 0. 047 7 pmol/L and 195 pmol/L, the proliferation rate of γδT cells increased remarkably （P 〈 0.05 ）. When the concentration was between 0. 191 pmol/L and 195 pmol/L, the cytotoxic activity of γδT cells to SW- 620 cells enhanced significantly （P 〈 0.05 ）. After induction with salvigenin for 72 h, expressions of GraB, PFP and CD107a were significantly higher （P 〈 0. 05 ）. When salvigenin concentration was between 0.763 pmol/L and 195 pmol/L, expression of p - ERK1/2 was significantly elevated. Conclusion At suitable concentrations, salvigenin can promote proliferation of human γδT cells and increase their cytotoxic activity against colon cancer cell SW - 620. A possible mechanism may be up - regulated expressions of PFP and GraB and activation of the p - ERK1/2 signaling pathway.