复方苦参对人末梢血γδT细胞杀伤胃癌细胞株SGC-7901作用的研究

目的探讨复方苦参对人γδT细胞杀伤胃癌细胞株SGC-7901的影响。方法用异戊烯焦磷酸法体外扩增人外周血γδT细胞,用不同浓度的复方苦参诱导γδT细胞SGC-7901细胞株24h,用MTT法检测复方苦参对这两种细胞生长抑制率的影响和LDH法检测γδT细胞的杀伤活性,用流式细胞术检测诱导前后的18T细胞和SGC-7901的凋亡率。结果γδT细胞培养10d时从扩增前4．21％增加到70．35％,CD44达94．0％。不同浓度的复方苦参对SGC-7901细胞株的抑制率（22．3％）均明显高于γδT细胞（-22．4％）,且γδT细胞的抑制率呈负的趋势,当复方苦参的浓度在1／50～1／400时γδT细胞负抑制率呈剂量依赖关系,且经复方苦参诱导24h的γδT细胞杀伤活性（83．6％）明显高于先诱导SGC-7901组（71．2％）,同时经复方苦参诱导24h的18T细胞凋亡率（4．64％）明显低于SGC-7901（49．23％）。结论复方苦参在临床常规使用浓度下,能够促进78T细胞的增殖,同时能够抑制肿瘤细胞的生长,且能够增强γδT细胞的杀伤活性,这一结果将有助于肿瘤的过继免疫治疗及为复方苦参应用于肿瘤治疗提供了临床依据。     

根皮素对人γδT细胞杀伤胃癌SGC-7901细胞的影响及机制探讨

目的:探讨根皮素对人γδT细胞杀伤胃癌SGC-7901细胞的影响及其机制。方法:IPP法扩增人外周血γδT细胞,不同浓度的根皮素作用于γδT细胞及胃癌SGC-7901细胞48小时后,MTT法检测γδT细胞及SGC-7901细胞的生长曲线,FCM检测γδT细胞PFP及GraB的表达;LDH释放法检测γδT细胞对SGC-7901细胞的杀伤活性;Western blot检测γδT细胞中Wnt3a的表达情况。结果:IPP作用10天后,γδT细胞比例由3.12%增加到79.6%。与对照组比较,浓度2.35~18.75μg/ml的根皮素作用后γδT细胞的增殖率显著提高(P<0.05),75μg/ml根皮素对SGC-7901细胞生长抑制率显著提高(P<0.05),且2.35~75μg/ml根皮素作用后的γδT细胞对SGC-7901细胞的杀伤活性明显增强(P<0.05),γδT细胞中PFP、GraB及Wnt3a的表达较对照组显著增加(P<0.05)。结论:根皮素能够增强γδT细胞对SGC-7901细胞的杀伤作用,其机制可能与根皮素促进γδT细胞的增殖,提高γδT细胞PFP、GraB表达及活化Wnt信号通路有关。     

水飞蓟宾增强人CIK细胞杀伤胃癌细胞株BCG-823的体外研究

为探讨水飞蓟宾对人CIK细胞杀伤胃癌细胞株BCG-823的影响及作用机制,分离健康志愿者外周血单个核细胞,体外诱导培养成人CIK细胞;收集培养扩增7d的CIK细胞,将其用不同浓度的水飞蓟宾诱导24h、48h、72h,CCK-8法检测其增殖情况;流式细胞术检测CIK细胞穿孔素、颗粒酶B、CD107a和IFN-γ的表达变化及水飞蓟宾对其凋亡的影响;LDH法检测CIK细胞对胃癌细胞株BCG-823的杀伤活性。结果显示,与对照组相比,浓度为(1.6~50)μg/mL的水飞蓟宾作用CIK细胞72h后,增殖率较对照组显著增加(P<0.05);穿孔素、颗粒酶B、CD107a和IFN-γ的表达以及活细胞率均不同程度增加(P<0.05);且对胃癌细胞BGC-823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CIK细胞有促增殖作用,且能增强其对胃癌细胞株BCG-823的杀伤活性。     

The effect of Fufangkushen on gastric cancer cell killing by lines SGC-7901 human γδT cell

Objective To explore the effect of Fufangkushen on gastric cancer cell killing by human γδT cells. Methods Isopentenyl pyrophosphate method was used to amplify human peripheral blood γδT cells in vitro. Fufangkushen at various concentrations was used to induce γδT cells and gastric cancer cell lines SGC-7901 for 24 hours, MTr assays was used to detect inhibitory effect of Fufangkushen on these cell lines, LDH assays was used to measure the cytotoxic activity of γδT cells, and flow cytometry was used to detect apoptosis of γδT cells and SGC-7901 before and after the treatment. Results Ten days after cultivation, proliferation ra-tio of γδT cells increased from 4.21% to 70.35% and CD44 was up to 94.0%. Inhibitory rate of Fufangkush-en on SGC-7901 at various concentrations was significantly higher than that on γδT cells (22.3% vs-22.4%, P<0.05). The negative inhibitory ratio on γδT cells showed a dose-dependent manner with Fufangkushen's concentrations ranging from 1/5 to 1/400. γδT cells cytotoxic activity to SGC-7901 induced by Fufangkushen for 24 h was higher than control group, (83.6% vs 71.2%, P<0.05). Apoptotic rate was significantly lower in γδT cells than in SGC-7901 (4.64% vs49.23%, P<0.05). Conclusion Fufangkushen, within routine concentration ranges, can promote γδT cells' proliferation, inhibit tumor cell growth and enhance γδT cells' cytotoxic activity. This may be beneficial to tumor adoptive immunotherapy and provide evidence for the appli-cation of Fufangkushen in the treatment of tumors.     

The effect of Fufangkushen on gastric cancer cell killing by lines SGC-7901 human γδT cell

Objective To explore the effect of Fufangkushen on gastric cancer cell killing by human γδT cells. Methods Isopentenyl pyrophosphate method was used to amplify human peripheral blood γδT cells in vitro. Fufangkushen at various concentrations was used to induce γδT cells and gastric cancer cell lines SGC-7901 for 24 hours, MTr assays was used to detect inhibitory effect of Fufangkushen on these cell lines, LDH assays was used to measure the cytotoxic activity of γδT cells, and flow cytometry was used to detect apoptosis of γδT cells and SGC-7901 before and after the treatment. Results Ten days after cultivation, proliferation ra-tio of γδT cells increased from 4.21% to 70.35% and CD44 was up to 94.0%. Inhibitory rate of Fufangkush-en on SGC-7901 at various concentrations was significantly higher than that on γδT cells (22.3% vs-22.4%, P<0.05). The negative inhibitory ratio on γδT cells showed a dose-dependent manner with Fufangkushen's concentrations ranging from 1/5 to 1/400. γδT cells cytotoxic activity to SGC-7901 induced by Fufangkushen for 24 h was higher than control group, (83.6% vs 71.2%, P<0.05). Apoptotic rate was significantly lower in γδT cells than in SGC-7901 (4.64% vs49.23%, P<0.05). Conclusion Fufangkushen, within routine concentration ranges, can promote γδT cells' proliferation, inhibit tumor cell growth and enhance γδT cells' cytotoxic activity. This may be beneficial to tumor adoptive immunotherapy and provide evidence for the appli-cation of Fufangkushen in the treatment of tumors.     

The effect of Fufangkushen on gastric cancer cell killing by lines SGC-7901 human γδT cell

Objective To explore the effect of Fufangkushen on gastric cancer cell killing by human γδT cells. Methods Isopentenyl pyrophosphate method was used to amplify human peripheral blood γδT cells in vitro. Fufangkushen at various concentrations was used to induce γδT cells and gastric cancer cell lines SGC-7901 for 24 hours, MTr assays was used to detect inhibitory effect of Fufangkushen on these cell lines, LDH assays was used to measure the cytotoxic activity of γδT cells, and flow cytometry was used to detect apoptosis of γδT cells and SGC-7901 before and after the treatment. Results Ten days after cultivation, proliferation ra-tio of γδT cells increased from 4.21% to 70.35% and CD44 was up to 94.0%. Inhibitory rate of Fufangkush-en on SGC-7901 at various concentrations was significantly higher than that on γδT cells (22.3% vs-22.4%, P<0.05). The negative inhibitory ratio on γδT cells showed a dose-dependent manner with Fufangkushen's concentrations ranging from 1/5 to 1/400. γδT cells cytotoxic activity to SGC-7901 induced by Fufangkushen for 24 h was higher than control group, (83.6% vs 71.2%, P<0.05). Apoptotic rate was significantly lower in γδT cells than in SGC-7901 (4.64% vs49.23%, P<0.05). Conclusion Fufangkushen, within routine concentration ranges, can promote γδT cells' proliferation, inhibit tumor cell growth and enhance γδT cells' cytotoxic activity. This may be beneficial to tumor adoptive immunotherapy and provide evidence for the appli-cation of Fufangkushen in the treatment of tumors.     

The effect of Fufangkushen on gastric cancer cell killing by lines SGC-7901 human γδT cell

Objective To explore the effect of Fufangkushen on gastric cancer cell killing by human γδT cells. Methods Isopentenyl pyrophosphate method was used to amplify human peripheral blood γδT cells in vitro. Fufangkushen at various concentrations was used to induce γδT cells and gastric cancer cell lines SGC-7901 for 24 hours, MTr assays was used to detect inhibitory effect of Fufangkushen on these cell lines, LDH assays was used to measure the cytotoxic activity of γδT cells, and flow cytometry was used to detect apoptosis of γδT cells and SGC-7901 before and after the treatment. Results Ten days after cultivation, proliferation ra-tio of γδT cells increased from 4.21% to 70.35% and CD44 was up to 94.0%. Inhibitory rate of Fufangkush-en on SGC-7901 at various concentrations was significantly higher than that on γδT cells (22.3% vs-22.4%, P<0.05). The negative inhibitory ratio on γδT cells showed a dose-dependent manner with Fufangkushen's concentrations ranging from 1/5 to 1/400. γδT cells cytotoxic activity to SGC-7901 induced by Fufangkushen for 24 h was higher than control group, (83.6% vs 71.2%, P<0.05). Apoptotic rate was significantly lower in γδT cells than in SGC-7901 (4.64% vs49.23%, P<0.05). Conclusion Fufangkushen, within routine concentration ranges, can promote γδT cells' proliferation, inhibit tumor cell growth and enhance γδT cells' cytotoxic activity. This may be beneficial to tumor adoptive immunotherapy and provide evidence for the appli-cation of Fufangkushen in the treatment of tumors.     

The effect of Fufangkushen on gastric cancer cell killing by lines SGC-7901 human γδT cell

Objective To explore the effect of Fufangkushen on gastric cancer cell killing by human γδT cells. Methods Isopentenyl pyrophosphate method was used to amplify human peripheral blood γδT cells in vitro. Fufangkushen at various concentrations was used to induce γδT cells and gastric cancer cell lines SGC-7901 for 24 hours, MTr assays was used to detect inhibitory effect of Fufangkushen on these cell lines, LDH assays was used to measure the cytotoxic activity of γδT cells, and flow cytometry was used to detect apoptosis of γδT cells and SGC-7901 before and after the treatment. Results Ten days after cultivation, proliferation ra-tio of γδT cells increased from 4.21% to 70.35% and CD44 was up to 94.0%. Inhibitory rate of Fufangkush-en on SGC-7901 at various concentrations was significantly higher than that on γδT cells (22.3% vs-22.4%, P<0.05). The negative inhibitory ratio on γδT cells showed a dose-dependent manner with Fufangkushen's concentrations ranging from 1/5 to 1/400. γδT cells cytotoxic activity to SGC-7901 induced by Fufangkushen for 24 h was higher than control group, (83.6% vs 71.2%, P<0.05). Apoptotic rate was significantly lower in γδT cells than in SGC-7901 (4.64% vs49.23%, P<0.05). Conclusion Fufangkushen, within routine concentration ranges, can promote γδT cells' proliferation, inhibit tumor cell growth and enhance γδT cells' cytotoxic activity. This may be beneficial to tumor adoptive immunotherapy and provide evidence for the appli-cation of Fufangkushen in the treatment of tumors.     

The effect of Fufangkushen on gastric cancer cell killing by lines SGC-7901 human γδT cell

Objective To explore the effect of Fufangkushen on gastric cancer cell killing by human γδT cells. Methods Isopentenyl pyrophosphate method was used to amplify human peripheral blood γδT cells in vitro. Fufangkushen at various concentrations was used to induce γδT cells and gastric cancer cell lines SGC-7901 for 24 hours, MTr assays was used to detect inhibitory effect of Fufangkushen on these cell lines, LDH assays was used to measure the cytotoxic activity of γδT cells, and flow cytometry was used to detect apoptosis of γδT cells and SGC-7901 before and after the treatment. Results Ten days after cultivation, proliferation ra-tio of γδT cells increased from 4.21% to 70.35% and CD44 was up to 94.0%. Inhibitory rate of Fufangkush-en on SGC-7901 at various concentrations was significantly higher than that on γδT cells (22.3% vs-22.4%, P<0.05). The negative inhibitory ratio on γδT cells showed a dose-dependent manner with Fufangkushen's concentrations ranging from 1/5 to 1/400. γδT cells cytotoxic activity to SGC-7901 induced by Fufangkushen for 24 h was higher than control group, (83.6% vs 71.2%, P<0.05). Apoptotic rate was significantly lower in γδT cells than in SGC-7901 (4.64% vs49.23%, P<0.05). Conclusion Fufangkushen, within routine concentration ranges, can promote γδT cells' proliferation, inhibit tumor cell growth and enhance γδT cells' cytotoxic activity. This may be beneficial to tumor adoptive immunotherapy and provide evidence for the appli-cation of Fufangkushen in the treatment of tumors.     

The effect of Fufangkushen on gastric cancer cell killing by lines SGC-7901 human γδT cell

Objective To explore the effect of Fufangkushen on gastric cancer cell killing by human γδT cells. Methods Isopentenyl pyrophosphate method was used to amplify human peripheral blood γδT cells in vitro. Fufangkushen at various concentrations was used to induce γδT cells and gastric cancer cell lines SGC-7901 for 24 hours, MTr assays was used to detect inhibitory effect of Fufangkushen on these cell lines, LDH assays was used to measure the cytotoxic activity of γδT cells, and flow cytometry was used to detect apoptosis of γδT cells and SGC-7901 before and after the treatment. Results Ten days after cultivation, proliferation ra-tio of γδT cells increased from 4.21% to 70.35% and CD44 was up to 94.0%. Inhibitory rate of Fufangkush-en on SGC-7901 at various concentrations was significantly higher than that on γδT cells (22.3% vs-22.4%, P<0.05). The negative inhibitory ratio on γδT cells showed a dose-dependent manner with Fufangkushen's concentrations ranging from 1/5 to 1/400. γδT cells cytotoxic activity to SGC-7901 induced by Fufangkushen for 24 h was higher than control group, (83.6% vs 71.2%, P<0.05). Apoptotic rate was significantly lower in γδT cells than in SGC-7901 (4.64% vs49.23%, P<0.05). Conclusion Fufangkushen, within routine concentration ranges, can promote γδT cells' proliferation, inhibit tumor cell growth and enhance γδT cells' cytotoxic activity. This may be beneficial to tumor adoptive immunotherapy and provide evidence for the appli-cation of Fufangkushen in the treatment of tumors.     

The effect of Fufangkushen on gastric cancer cell killing by lines SGC-7901 human γδT cell

Objective To explore the effect of Fufangkushen on gastric cancer cell killing by human γδT cells. Methods Isopentenyl pyrophosphate method was used to amplify human peripheral blood γδT cells in vitro. Fufangkushen at various concentrations was used to induce γδT cells and gastric cancer cell lines SGC-7901 for 24 hours, MTr assays was used to detect inhibitory effect of Fufangkushen on these cell lines, LDH assays was used to measure the cytotoxic activity of γδT cells, and flow cytometry was used to detect apoptosis of γδT cells and SGC-7901 before and after the treatment. Results Ten days after cultivation, proliferation ra-tio of γδT cells increased from 4.21% to 70.35% and CD44 was up to 94.0%. Inhibitory rate of Fufangkush-en on SGC-7901 at various concentrations was significantly higher than that on γδT cells (22.3% vs-22.4%, P<0.05). The negative inhibitory ratio on γδT cells showed a dose-dependent manner with Fufangkushen's concentrations ranging from 1/5 to 1/400. γδT cells cytotoxic activity to SGC-7901 induced by Fufangkushen for 24 h was higher than control group, (83.6% vs 71.2%, P<0.05). Apoptotic rate was significantly lower in γδT cells than in SGC-7901 (4.64% vs49.23%, P<0.05). Conclusion Fufangkushen, within routine concentration ranges, can promote γδT cells' proliferation, inhibit tumor cell growth and enhance γδT cells' cytotoxic activity. This may be beneficial to tumor adoptive immunotherapy and provide evidence for the appli-cation of Fufangkushen in the treatment of tumors.     

The effect of Fufangkushen on gastric cancer cell killing by lines SGC-7901 human γδT cell

Objective To explore the effect of Fufangkushen on gastric cancer cell killing by human γδT cells. Methods Isopentenyl pyrophosphate method was used to amplify human peripheral blood γδT cells in vitro. Fufangkushen at various concentrations was used to induce γδT cells and gastric cancer cell lines SGC-7901 for 24 hours, MTr assays was used to detect inhibitory effect of Fufangkushen on these cell lines, LDH assays was used to measure the cytotoxic activity of γδT cells, and flow cytometry was used to detect apoptosis of γδT cells and SGC-7901 before and after the treatment. Results Ten days after cultivation, proliferation ra-tio of γδT cells increased from 4.21% to 70.35% and CD44 was up to 94.0%. Inhibitory rate of Fufangkush-en on SGC-7901 at various concentrations was significantly higher than that on γδT cells (22.3% vs-22.4%, P<0.05). The negative inhibitory ratio on γδT cells showed a dose-dependent manner with Fufangkushen's concentrations ranging from 1/5 to 1/400. γδT cells cytotoxic activity to SGC-7901 induced by Fufangkushen for 24 h was higher than control group, (83.6% vs 71.2%, P<0.05). Apoptotic rate was significantly lower in γδT cells than in SGC-7901 (4.64% vs49.23%, P<0.05). Conclusion Fufangkushen, within routine concentration ranges, can promote γδT cells' proliferation, inhibit tumor cell growth and enhance γδT cells' cytotoxic activity. This may be beneficial to tumor adoptive immunotherapy and provide evidence for the appli-cation of Fufangkushen in the treatment of tumors.     

尼美舒利提高γδT细胞杀伤胃癌细胞的机制探讨

目的:探讨尼美舒利对人γδT细胞功能影响。方法:按常规方法培养γδT细胞,收集培养第9天的γδT细胞用不同浓度的尼美舒利(分别为0.25、0.5、1、2、4μmol/L)诱导24 h后收集培养上清液检测细胞因子IFN-γ、TNF-α、IL-12,沉淀细胞流式细胞测定穿孔素、粒酶B和NKG2D,LDH法检测γδT细胞杀伤胃癌细胞活性。结果:经尼美舒利诱导后γδT细胞穿孔素、粒酶B表达量(分别为62.8%和72.7%)明显高于对照组(分别为51.4%和60.9%)P0.05,尤其尼美舒利浓度在1μmol/L时最显著;经尼美舒利诱导后γδT细胞分泌IFN-γ和TNF-α浓度(分别为262.3 ng/L和177.5 ng/L)明显高于对照组(分别为196.1 ng/L和158.5 ng/L)P0.05;分泌的IL-12浓度与对照组比较无明显差异(P0.05)。γδT细胞杀伤胃癌细胞SGC-7901和BCG-823的活性在1μmol/L时最高(分别为73%和70%),明显高于对照组(分别为54%和53%)P0.05。结论:经尼美舒利诱导后γδT细胞的穿孔素、粒酶B含量和γδT细胞杀伤胃癌细胞SGC-7901、BCG-823活性明显高于对照组。其培养上清液中IFN-γ和TNF-α浓度也明显高于对照组。这一结果为临床尼美舒利预防和治疗消化道肿瘤提供了实验依据。     

MiR-141对胃癌细胞SGC-7901增殖、侵袭能力的影响

目的观察miR-141在胃癌组织中的表达变化,及miR-141对胃癌细胞株SGC-7901增殖、侵袭能力的影响。方法收集116例胃癌患者的胃癌组织及癌旁正常胃组织,采用real-time PCR方法检测miR-141的表达水平,将化学合成的miR-141拟物和miR-141抑制剂转染至胃癌SGC-7901细胞,分别于培养12、24、48、72 h后采用MTT法检测细胞增殖能力,培养24 h后Transwell小室检测细胞侵袭能力,采用Western blot法检测细胞中的MMP-2、MMP-9蛋白的表达。结果胃癌组织中miR-141相对表达量显著低于癌旁组织(P0.01)。转染miR-141模拟物24、48、72 h后,SGC-7901细胞增殖能力显著降低,转染miR-141抑制剂后SGC-7901细胞增殖能力显著升高(P0.01)。转染miR-141模拟物24 h后,穿膜细胞数、基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)蛋白相对表达量显著降低,转染miR-141抑制剂后穿膜细胞数、MMP-2、MMP-9蛋白相对表达量显著升高(P0.01)。结论胃癌组织中miR-141低表达可能与胃癌的发生发展有关,miR-141抑制胃癌细胞增殖、侵袭的作用机制可能与下调MMP-2、MMP-9蛋白表达有关。     

γδT细胞的抗原识别机制

Ｔ细胞表面表达两类抗原受体（ＴＣＲ）：ＴＣＲαβ和ＴＣＲγδ。ＴＣＲαβ可特异地识别由抗原呈递细胞（ＡＰＣ）表面Ⅰ类或Ⅱ类ＭＨＣ分子呈递的抗原肽，而ＴＣＲγδ则主要以ＭＨＣ非限制方式识别各类抗原。最近对ＴＣＲγδ所识别的抗原类型及方式进行了较为深入的研究，本文就其进展作一述评。１　ＴＣＲγδ的多样性和分布特点提示其抗原识别的特殊性　　同ＴＣＲαβ和免疫球蛋白（Ｉｇ）类似，ＴＣＲγδ基因由重组的Ｖ、Ｄ、Ｊ和Ｃ区组成。虽然γ、δ位点的Ｖ区多样性不及α和β，但其连接区多样性则使ＴＣＲγδ存在甚至超过Ｔ…     

siRNA沉默胃癌SGC-7901细胞HO1基因的细胞系的建立及其生物学行为

目的 构建HO1基因的真核干扰表达载体,评估其转染人胃癌细胞系SGC-7901细胞后对HO1基因的干扰效果及其功能.方法 将外源性重组真核干扰表达载体HO1基因(pS/HO1)转染到人胃癌细胞系SGC-7901内,经G418筛选并建立siRNA表达载体稳定沉默胃癌SGC-7901细胞HO1基因的细胞系,分为SGC-7901-pS/HO1组,转染空质粒细胞(SGC-7901-pS)组和未处理细胞(SGC-7901)组;用实时荧光定量PCR和蛋白印迹验证HO1基因在各组细胞中的表达,并通过CCK-8和克隆形成实验分别观察HO1基因被干扰后细胞的生物学行为.结果 与SGC-7901-pS组相比,SGC-7901-pS/HO1细胞中HO1基因蛋白表达明显减少,降低了5.58倍(0.321±0.051 vs 1.675±0.153,P＜0.05);与对照组相比较,SGC-7901-pS/HO1实验组较SGC-7901-pS对照组细胞增殖数量明显减少(P＜0.001);与转染pS空载体的SGC-7901-pS细胞对照组相比,SGC-7901-pS/HO1细胞的克隆形成明显减少,降低了3.45倍(8.32±1.142 vs 2.32 ±0.362,P＜0.05).结论 HO1基因真核siRNA表达载体筛选成功,为继续深入的研究HO1基因在胃癌中的功能提供了依据.     

乙酰紫草素注射液对人胃癌细胞SGC-7901的抑制作用

目的:研究乙酰紫草素注射液的体外抗人胃癌细胞SGC-7901肿瘤作用。方法:以不同浓度的乙酰紫草素注射液作用于SGC-7901细胞,分别用MTT法、细胞生长曲线研究其对SGC-7901的增殖抑制作用。结果:MTT显示乙酰紫草素注射液浓度在0.1-3.2mg·L-1时,对SGC-7901细胞的抑制率为22-90%,IC50值为0.428±0.07mg.L-1。生长曲线提示乙酰紫草素注射液对SGC-7901细胞的抑制作用呈明显时效和量效关系。结论:乙酰紫草素注射液对人胃癌细胞SGC-7901有较强的增殖抑制作用。     

辣椒素对人γδT细胞杀伤人骨肉瘤HOS细胞的影响

探讨辣椒素对人γδT细胞体外增殖及杀伤骨肉瘤细胞的影响及作用机制。分离健康人PBMC,加入到含异戊烯焦磷酸(isopentenyl pyrophosphate,IPP)和IL-2的DMEM-F12完全培养基中诱导培养γδT细胞。不同浓度的辣椒素作用于γδT细胞48h后,CCK-8法检测各药物浓度组γδT细胞的增殖能力,采用流式细胞术检测各组γδT细胞穿孔素、颗粒酶B、CD107a及IFN-γ的表达,用LDH释放法检测各组γδT细胞对骨肉瘤HOS细胞的体外杀伤活性。结果显示,培养7d时各组γδT细胞纯度达到了(85.33±6.07)%。浓度为3.2~50μmol/L的辣椒素能显著促进γδT细胞的增殖,在浓度为25μmol/L时其穿孔素、颗粒酶B、CD107a及IFN-γ的表达均显著高于对照组,且对人骨肉瘤HOS细胞的体外杀伤活性显著增强。以上结果提示辣椒素在体外能通过促进γδT细胞穿孔素、颗粒酶B和IFN-γ的表达上调显著增强其抗骨肉瘤细胞活性。     

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

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-1l16细胞活性的影响。Western blot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 ～3.125μmol/L时对γδT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0.1～10 μmol/L白藜芦醇作用的γδT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γδT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。