IL-18基因修饰对肺癌细胞来源exosome诱导杀伤肿瘤细胞作用的影响  

Effect of interleukin 18 gene modification on anti-tumor activity induced by lung cancer cell-derived exosomes

作　　者：张在云[1] 李晓梅[1] 王涓冬[2] 孙建华[1] 姜玉华[1] 潘祥林[2] 

机构地区：[1]山东大学第二医院肿瘤防治中心,山东济南250033 [2]山东大学第二医院血液内科,山东济南250033

出　　处：《中国病理生理杂志》2011年第9期1758-1761,共4页Chinese Journal of Pathophysiology

基　　金：山东省自然科学基金资助项目(No.Y2007C111)

摘　　要：目的:研究细胞介素18(IL-18)基因修饰对肺癌细胞来源exosome诱导杀伤肿瘤细胞作用的影响,以探讨高效exosome疫苗的制备。方法:提取IL-18基因修饰的NCI-H460细胞(IL-18/H460)、pcDNA3.1+载体修饰细胞(DNA3.1/H460)及未修饰NCI-H460细胞(NCI-H460)上清液的exosome,透射电镜下观察exo-some形态;用Western blotting方法检测exosome中热休克蛋白70(HSP70)、人白细胞抗原(HLA)及IL-18的表达;用exosome直接刺激活化T细胞,用乳酸脱氢酶(LDH)法测定T细胞对NCI-H460细胞的杀伤作用;用exosome冲击树突状细胞(DCs),然后活化T细胞,测定对NCI-H460细胞的杀伤作用,计算杀伤率。结果:透射电镜下观察exosome具有典型的形态;3组exosome中均有HSP70及HLA蛋白表达,IL-18/H460组有IL-18蛋白表达。效靶比25∶1、10∶1、5∶1时exosome直接提高活化T细胞的杀伤率,IL-18/H460组为(38.45±5.42)%、(25.17±3.94)%和(11.75±3.22)%,exosome冲击DCs活化T细胞的杀伤率,IL-18/H460组为(89.05±4.06)%、(64.97±6.02)%和(40.16±4.98)%,均高于其余2组。Exosome冲击DCs活化T细胞的杀伤作用强于exosome直接刺激活化的T细胞。结论:IL-18基因修饰能增强NCI-H460细胞来源exosome诱导的杀伤肿瘤细胞作用。AIM： To study the effect of interleukin 18（IL-8） gene modification on anti-tumor activity induced by lung cancer cell-derived exosomes.METHODS： Exosomes isolated from the supernatants of IL-18 gene-modified NCI-H460 lung cancer cells（IL-18/H460）,pcDNA3.1＋ vector-modified cancer cells（DNA3.1/H460） and non-modified NCI-H460 lung cancer cells（NCI-H460） were observed under transmission electron microscope.The expression of heat-shock protein 70（HSP70）,human leukocyte antigen（HLA） and IL-18 were determined by Western blotting.T lymphocytes were activated by exosomes or exosome-pulsed dendritic cells（DCs）.The activity of T cells for killing lung cancer cells were detected by lactate dehydrogenase（LDH） method.The killing rates were calculated and compared.RESULTS： Exosomes showed typical morphous under transmission electron microscope.The protein levels of HSP70 and HLA were detected in the exosomes of all 3 groups,and IL-18 protein was only observed in IL-18/H460 group.The killing rates of exosome-activated T cells in IL-18/H460 group with the ratio of effector cell to target cell at 25∶1,10∶1 and 5∶1 were（38.45±5.42）%,（25.17±3.94）% and（11.75±3.22）%,respectively.The killing rates of exosome-pulsed DC-activated T cells in this group were（89.05±4.06）%,（64.97±6.02）% and（40.16±4.98）%,respectively.The killing rates in IL-18/H460 group were higher than those in DNA3.1/H460 group and NCI-H460 group.The anti-tumor efficacy of exosome-pulsed DC-activated T cells was stronger than that of exosome-activated T cells.CONCLUSION： IL-18 gene modification enhances the anti-tumor activity induced by NCI-H460 lung cancer cell-derived exosomes.

关 键 词：白细胞介素18 基因修饰 肺肿瘤 EXOSOME 癌症疫苗 

分 类 号：R363[医药卫生—病理学]