IL-15转染人CIK细胞对HT-29的杀瘤效果探究  

Tumor-killing effect of IL-15 transfected human CIK cells on HT-29 cells

作　　者：王柏涛 梁宇堃 王丙萍 杨超旗 高艳伟 WANG Bai-tao;LIANG Yu-kun;WANG Bing-ping;YANG Chao-qi;GAO Yan-wei(Endoscopy Center,Inner Mongolia Autonomous Region People’s Hospital,Hohhot 010017,Inner Mongolia,China;Abdominal Tumer Surgery,Inner Mongolia Autonomous Region People’s Hospital,Hohhot 010017,Inner Mongolia,China;Department of Emergency Medicine,Ulanqab Center Hospital,Ulanqab 012000,Inner Mongolia,China;Inner Mongolia Cancer Research Institute,Hohhot 010017,Inner Mongolia,China)

机构地区：[1]内蒙古自治区人民医院内镜中心,内蒙古呼和浩特010017 [2]内蒙古自治区人民医院腹部肿瘤外科,内蒙古呼和浩特010017 [3]乌兰察布市中心医院急救医学科,内蒙古乌兰察布012000 [4]内蒙古自治区肿瘤研究所,内蒙古呼和浩特010017

出　　处：《生物医学工程与临床》2023年第5期650-655,共6页Biomedical Engineering and Clinical Medicine

摘　　要：目的探讨白细胞介素15(IL-15)基因(IL-15)转染人细胞因子诱导杀伤(CIK)细胞对HT-29[人结肠癌(CRC)细胞株]的杀瘤效应。方法从血液样本制备CIK细胞后,用含IL-15的慢病毒感染CIK细胞,制备转基因CIK细胞(IL-15-CIK细胞)。将IL-15-CIK细胞、CIK细胞分别与HT-29细胞共培养,按效靶比5︰1、10︰1、20︰1、25∶1培养14 d。分别评估IL-15-CIK细胞、CIK细胞对HT-29细胞的IL-15、干扰素γ(IFN-γ)、肿瘤坏死因子α(TNF-α)含量变化和杀瘤效应。结果成功制备CIK细胞及IL-15-CIK细胞。效靶比25∶1,IL-15-CIK细胞与CIK细胞对CRC细胞的杀伤最高(72.56%±5.66%、52.33%±3.46%),且两种细胞差异有统计学意义(P<0.05)。即IL-15-CIK细胞对HT-29细胞的杀伤能力强于CIK细胞。在IL-15-CIK细胞及CIK细胞培养7 d、11 d、14 d中,两种细胞上清液中IL-15、IFN-γ及TNF-α的含量均随着时间的增加而升高,杀瘤后任何两组间比较,差异有统计学意义(P<0.05);但在IL-15-CIK细胞中观察到,TNF-α含量在11 d与14 d内差异无统计学意义(P>0.05),其余时间相比,差异均有统计学意义(P<0.05)。IL-15-CIK细胞及CIK细胞以E∶T=25∶1对HT-29细胞株作用后,检测到两种细胞上清液中IL-15、TNF-α、IFN-γ含量较作用前含量增加[IL-15-CIK细胞:IL-15(45.37±2.15)pg/mL vs(35.49±1.09)pg/mL,TNF-α(34.83±1.63)pg/mL vs(28.04±0.41)pg/mL,IFN-γ(42.34±1.50)pg/mL vs(26.08±0.57)pg/mL。CIK细胞:IL-15(21.18±0.76)pg/mL vs(20.34±1.07)pg/mL,TNF-α(20.04±3.03)pg/mL vs(18.04±0.27)pg/mL,IFN-γ(23.90±1.33)pg/mL vs(19.06±0.34)pg/mL]。结论IL-15-CIK细胞对CRC细胞株的杀伤能力较CIK细胞强,应用IL-15转染人CIK细胞可增强CIK细胞杀瘤效应。Objective To investigate the tumor-killing effect of interleukin-15(IL-15)gene(IL-15)transfected human cytokine-induced killer(CIK)cells on HT-29(human colorectal cancer cell line).Methods After CIK cells were prepared from blood samples,transgenic CIK cells(IL-15-CIK cells)were prepared by infecting CIK cells with lentivirus containing IL-15.The IL-15-CIK cells and CIK cells were co-cultured with HT-29 cells,respectively,and cultured at effector-target ratios of 5:1,10:1,20:1,and 25:1 for 14-day.The changes of IL-15,interferonγ(IFN-γ),tumor necrosis factor-α(TNF-α)content and tumor-killing effect of IL-15-CIK cells and CIK cells on HT-29 cells were evaluated respectively.Results The CIK cells and IL-15-CIK cells were successfully prepared.The effector-to-target ratio was 25:1,IL-15-CIK cells and CIK cells showed the highest tumor-killing effect on CRC cells(72.56%±5.66%,52.33%±3.46%),and the difference was statistically significant between 2 cells(P<0.05),the tumor-killing effect of IL-15-CIK cells to HT-29 cells was stronger than that of CIK cells.The contents of IL-15,IFN-γand TNF-αin the supernatant of IL-15-CIK cells and CIK cells cultured for 7-day,11-day and 14-day increased with time,the difference was statistically significant between paired groups after tumor-killing(P<0.05),while there was no significant difference of TNF-αcontent in IL-15-CIK cells at 11-day and 14-day(P>0.05),and the differences were statistically significant at other time(P<0.05).After IL-15-CIK cells and CIK cells were treated with E:T=25:1 on HT-29 cell line,the contents of IL-15,TNF-αand IFN-γin supernatant of 2 cells were higher than those before tumor-killing[IL-15-CIK cell:IL-15(45.37±2.15)pg/mL vs(35.49±1.09)pg/mL,TNF-α(34.83±1.63)pg/mL vs(28.04±0.41)pg/mL,IFN-γ(42.34±1.50)pg/mL vs(26.08±0.57)pg/mL.CIK cells:IL-15(21.18±0.76)pg/mL vs(20.34±1.07)pg/mL,TNF-α(20.04±3.03)pg/mL vs(18.04±0.27)pg/mL,IFN-γ(23.90±1.33)pg/mL vs(19.06±0.34)pg/mL].Conclusion It is demonstrated that the killing effect of IL-15-C

关 键 词：白细胞介素15(IL-15) 白细胞介素15(IL-15)基因 CIK细胞 杀瘤效应 

分 类 号：R73-36[医药卫生—肿瘤]