水飞蓟宾对γδT细胞杀伤胃癌细胞SGC-7901的影响及其机制初探

探讨水飞蓟宾对人γδT细胞杀伤胃癌细胞SGC-7901的影响及其作用机制。分离健康志愿者外周血单个核细胞,体外经多种细胞因子诱导培养为γδT细胞;收集培养、扩增7d后的γδT细胞,将其用不同浓度的水飞蓟宾诱导24h、48h及72h,CCK-8法检测γδT细胞增殖情况;流式细胞术(FCM)检测γδT穿孔素(perforin,PFP)、细胞颗粒酶B(Granzyme B,Gran B)及CDl07a的表达;western-blot法检测γδT细胞P-ERK1/2、Bcl-2、P-AKT、β-catenin的表达;乳酸脱氢酶(LDH)法检测γδT细胞对胃癌细胞SGC-7901的杀伤活性。结果发现浓度在1.6~100μg/ml的水飞蓟宾作用γδT细胞72h后,γδT细胞增殖率较对照组显著增加(P0.05),且在25μg/ml时达到最高峰;将6.25~100μg/ml水飞蓟宾诱导γδT细胞72h后,γδT细胞的PFP、Gran B及CDl07a的表达以及P-ERK1/2、Bcl-2、P-AKT、β-catenin的表达与对照组比较均不同程度增加(P0.05),且对胃癌细胞SGC-7901的杀伤活性显著增强(P0.05)。以上实验结果提示一定浓度的水飞蓟宾对γδT细胞具有促进增殖作用,其机制可能与激活P-ERK1/2、Bcl-2、P-AKT及β-catenin信号通路有关。一定浓度的水飞蓟宾能够增加γδT细胞对胃癌细胞的杀伤活性,其作用机制可能与水飞蓟宾能够增加γδT细胞的穿孔素、Gran B及CDl07a的表达有关。     

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.     

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。     

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。     

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。     

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。     

补中益气汤对SGC-7901人胃癌细胞株酶活性影响的研究

目的研究补中益气汤对人胃癌细胞株酶活性影响,探讨该药作用机制。方法以人胃腺癌细胞株SGC-7901为受试对象,利用血清药理学和酶细胞化学染色法,观察SGC-7901人胃腺癌细胞株的酶活性的改2+变。结果酶细胞化学法显示,中药组细胞质膜标志酶Mg-ATP酶和内质网标志酶G-6-P酶的酶反应颗粒小,数量减少,阳性细胞两种酶的表达量明显低于对照组(<0.05)。结论补中益气汤抗肿瘤作用可能与其降低2+胃癌细胞株Mg-ATP酶和G-6-P酶的活性相关。     

γδT细胞在疾病中的作用

γδT细胞是皮肤表皮内淋巴细胞和粘膜组织上皮内淋巴细胞的主要成分之一,为非特异性免疫细胞。活化的γδT细胞具有较强的杀伤活性,具有抗感染、抗肿瘤作用;γδT细胞还可以维持免疫耐受,调节免疫应签,异常可导致自身免疫性疾病的发生。因此尽管γδT细胞在人类庞大复杂的免疫体系中数量较少,但却具有不可替代的重要功能。     

活血行气方抑制人胃癌SGC-7901细胞株增殖及诱导其凋亡的实验研究

目的探讨活血行气方药物血清治疗人胃癌SGC-7901的作用。方法采用血清药理学方法,制备药物血清,应用MTT法测定细胞增殖抑制率,在光、电镜下观察细胞凋亡的的形态学变化。结果药物血清作用24h,中药小剂量组(5μl/ml)未见明显抑制作用,其余各组(10μl/ml,20μl/ml)均表现出不同程度的抑制作用;作用48h,中药小剂量组出现抑制作用,各药物血清组随着作用时间的延长抑制更明显,抑制效应呈时间依赖性。顺铂的抑制作用强于药物血清。光镜下,HE染色药物血清组细胞,为圆形或卵圆形,核染色质密集。电镜下人胃癌SGC-7901细胞为圆形,微绒毛及突起明显减少,伪足消失,线粒体空泡化,核染色质凝集,可见调亡小体。凋亡细胞百分率与药物浓度呈正相关。结论活血行气方具有明显抑制人胃癌SGC-7901细胞增殖、诱导细胞凋亡的作用,活血行气方与顺铂合用有协同作用。     

Gardiquimod增强人γδT细胞杀伤肿瘤细胞的研究

目的研究Toll样受体7激动剂(Gardiquimod)对人γδT细胞杀伤肿瘤作用的影响。方法以异戊烯焦磷酸法扩增人外周血γδT细胞。用不同浓度的Gardiquimod处理γδT细胞和肺癌细胞A549,MTT法检测细胞增殖情况。LDH法检测Gardiquimod处理后γδT细胞对A549的杀伤活性;流式细胞术检测处理前后γδT细胞上CD107a、穿孔素和颗粒酶的表达情况。结果 Gardiquimod在5.0~0.31μg/ml浓度可明显促进γδT细胞增殖,同时抑制A549增殖。Gardiquimod处理γδT细胞后,杀伤A549能力明显增强,且CD107a、颗粒酶及穿孔素的表达显著升高,与对照组相比,有统计学差异(P<0.05)。结论 Gardiquimod通过增高γδT细胞上的CD107a、颗粒酶及穿孔素的表达来增强其杀伤肿瘤作用。     

白桦酯酸对人CIK细胞杀伤胃癌细胞株SGC-7901的功能影响及机制研究

目的 观察白桦酯酸作用前后对人CIK细胞增殖及对胃癌SGC-7901细胞杀伤活性的影响,探讨其发生的机制.方法 分离健康者外周血单个核细胞( PBMC)在体外经多种细胞因子诱导为CIK细胞,收集培养第10天的CIK细胞,给予不同浓度白桦酯酸诱导48 h,四甲基偶氮唑蓝(MTT)法检测人CIK细胞增殖率;流式细胞术(FCM)检测白桦酯酸作用前后CIK细胞穿孔素(PFP)、颗粒酶B(GrB)、CD107a的表达变化;乳酸脱氢酶(LDH)释放法测定白桦酯酸对CIK细胞杀伤胃癌细胞株SGC-7901的活性影响;Western blot检测药物诱导前后CIK细胞胞外信号调节激酶(ERK1/2)和接头蛋白76KD含有SH2结构域的白细胞特异性磷酸蛋白(SLP-76)、T细胞活化连接分子(LAT)的表达变化.结果 白桦酯酸浓度在0.08 ～ 10μg/ml时能促进CIK细胞生长;经白桦酯酸诱导后的CIK细胞PFP、GrB、CD107a的表达显著高于对照组(P＜0.05),对胃癌SGC-7901细胞的杀伤活性亦显著高于对照组(P＜0.05);经白桦酯酸作用的CIK细胞,其SLP-76、LAT、ERK1/2的表达不同程度增加,显著高于对照组(P＜0.05).结论 白桦酯酸在一定浓度范围内能够促进CIK细胞增殖,并增强其对胃癌SGC-7901细胞的杀伤活性,其机制可能与活化SLP-76、LAT、ERK1/2,上调CIK细胞表面PFP、GrB、CD107a的表达有关.