丰富环境对肺癌荷瘤小鼠肿瘤组织NK细胞活性、Ki67蛋白表达的影响

摘要

Objective:To investigate the effects of enriched environment(EE)on the activity of NK cells and tumor tissue of mouse lung cancer Ki67 protein expression,so as to provide experimental basis for clinical application of enriched environment intervention in lung cancer.Methods:24 mice in open field test to screen the active medium group were randomly divided into the standard environment(SE)model group and EE model group,12 mice in each group.Mice LLC lung cancer cells were cultured to logarithmic growth phase under normal conditions,and were inoculated within the right arms of the mice.Normal environment of lung cancer model was established in the standard environment model group.Enriched environment model was established in the environment of lung cancer model group.At the end of four weeks,two groups were sacrificed.The activity of NK cells was detected by MTT.Observing the pathology morphology of the two groups by HE staining.Immunohistochemistry was used to detect the expression of Ki67 protein of the tumor tissue.Results:The model of LLC lung cancer was successfully established,and tumor formation rate was 100%.The NK cells activity tested by MTT:The EE group was(30.02±5.79)%,and the SE group was(25.49±4.51)%.The EE group had higher NK cell activity compared to SE group(P<0.05).The results of HE staining showed poor differentiation in EE and SE group.The positive expression of Ki67 protein in EE and SE group were 75.0% and 91.7%,respectively.The SE group had higher Ki67 protein expression compared to EE group(P<0.05).Conclusion:Enriched environment enhanced the natural killer cell activity,and reduced the Ki67 expression,therefore influenced the immune function.%目的:探讨丰富环境对肺癌荷瘤小鼠肿瘤组织NK细胞活性、Ki67蛋白表达的影响,为丰富环境干预肺癌临床推广应用提供实验依据.方法:动物分组:将旷场试验筛选出的中等活跃组小鼠24只,随机分为标准环境模型组和丰富环境模型组,每组12只.常规条件下培养小鼠LLC肺癌细胞至对数生长期,无菌接种于小鼠右侧腋下,建立小鼠肺癌模型.标准环境模型组为建肺癌模型后正常环境中饲养.丰富环境模型组为建肺癌模型后丰富环境中饲养.4周后取材.MTT法检测小鼠NK细胞活性.HE染色观察两组肿瘤病理学形态.免疫组织化学SP法检测肿瘤组织中Ki67的表达.结果:小鼠LLC肺癌模型成瘤率100%.丰富环境组小鼠NK细胞活性为(30.02±5.79)%,标准环境组小鼠NK细胞活性为(25.49±4.51)%,丰富环境组小鼠NK细胞活性明显大于标准环境模型组小鼠,差异有统计学意义(P<0.05).丰富环境组和标准环境组小鼠均为低分化的恶性肿瘤.丰富环境组和标准环境组小鼠肿瘤组织Ki67阳性表达率分别为75.0%、91.7%,差异有统计学意义.结论:丰富环境可能通过增强NK细胞活性,调节肿瘤组织Ki67蛋白表达,减少肿瘤发生、发展.