文献类型：期刊文献

中文题名：荧光素酶标记的Lewis肺癌细胞转移前微环境小鼠模型建立与评价

英文题名：Establishment and evaluation of pre-metastatic niche mouse model labelled with luciferase in lewis lung cancer cells

作者：明海霞[1,2,3,4];申明[1,2,3,4];陈彦文[1];李杨[1,2,3,4];杨玲玲[1];梁乾坤[1];刘兆华[1,2,3,4]

第一作者：明海霞

机构：[1]甘肃中医药大学基础医学院,甘肃兰州730000;[2]甘肃中医药大学中西医结合基础学科,甘肃兰州730000;[3]甘肃省高校重大疾病分子医学与中医药防治研究重点实验室,甘肃兰州730000;[4]甘肃省中医药防治慢性疾病重点实验室,甘肃兰州730000

第一机构：甘肃中医药大学基础医学院（敦煌医学研究所）

年份：2023

卷号：39

期号：12

起止页码：1083

中文期刊名：细胞与分子免疫学杂志

外文期刊名：Chinese Journal of Cellular and Molecular Immunology

收录：CSTPCD;;北大核心:【北大核心2020】；CSCD:【CSCD_E2023_2024】；PubMed;

基金：甘肃省“双一流”科研重点项目(GSSYLXM-05);甘肃省科技厅自然科学基金项目(20JR5RA178,20JR10RA320);甘肃省中医药研究中心开放课题(zyzx-2020-zx2);甘肃中医药大学科学研究与创新基金项目(2020KCZD-4,2021KCZD-1);甘肃中医药大学中西医结合基础学科科研培育项目(2020-2021-12);兰州市科技计划项目(2020-ZD-55,2021-1-96);甘肃省教育科技创新项目(2021CXZX-745)。

语种：中文

中文关键词：肺癌;转移前微环境;肿瘤转移;动物模型建立

外文关键词：lung cancer;the pre-metastatic niche;tumor metastasis;animal model

摘要：目的 通过选择荧光素酶标记的Lewis(Luc-Lewis)肺癌细胞建立肺转移前微环境小鼠模型,采用定性、定量的方法评价该模型是否成功。方法 C57BL/6小鼠分为正常对照组和模型组,每组15只,模型组小鼠通过尾静脉注射Luc-Lewis肺癌细胞建立转移前微环境模型。每日记录各组小鼠体质量,运用高通量酶标仪检测小鼠体内肿瘤荧光信号。取小鼠肺组织观察转移情况,HE染色观察肺组织病变情况,实时定量PCR和Western blot法分别检测转移前肺组织微环境形成的特异性标志物赖氨酰氧化酶(LOX)、基质金属蛋白酶9(MMP9)、多功能蛋白聚糖(VCAN)、纤连蛋白(FN)的mRNA和蛋白表达。结果 与正常组相比,模型组小鼠体质量显著下降,精神萎靡。模型组小鼠肺脏可检测到明确的荧光信号,且信号强度与造模天数呈正相关;14 d时模型组小鼠LOX、 MMP9、 VCAN、 FN的mRNA和蛋白表达水平显著升高;14 d时模型组小鼠肺组织肺间隙增厚,肺泡萎缩,大量炎性细胞浸润,第21天时可观察到模型组小鼠肺组织已形成转移病灶。通过以上实验结果提示转移前微环境成熟时间约为14 d。结论 成功建立了Lewis肺癌肿瘤转移前微环境小鼠模型。
Objective This study aimed to establish a pre-metastatic niche mouse model utilizing luciferase-labeled Lewis(Luc-Lewis)lung cancer cells and to assess the efficacy of this model employing both qualitative and quantitative methods.Methods C57BL/6 mice were categorized into two groups:a normal control group and a model group,each containing 15 individual mice.The pre-metastatic niche model was established via tail vein injection of Luc-Lewis lung cancer cells.Body mass were measured daily for all groups.Tumor fluorescence signals within the mice were detected using a high-throughput enzyme marker instrument.Lung tissue specimens were harvested to evaluate metastatic progression.HE staining was used to assess histopathological changes.Real-time quantitative PCR and Western blot analysis were used to detect the mRNA and protein expression of lysyl oxidase(LOX),matrix metalloproteinase 9(MMP9),versican(VCAN),and fibronectin(FN),which are the specific markers for the formation of the microenvironment of lung tissues before metastasis.Results Significant declines in body mass and observable lethargy were noted in the model group when compared to the control group.Distinct fluorescence signals were observed in the lung tissue of the model group,demonstrating a positive correlation with the duration of model establishment.By day 14,elevated mRNA and protein expression levels of LOX,MMP9,VCAN,and FN were significantly evident.In addition,histopathological evaluations revealed augmented interstitial thickness,alveolar atrophy and significant inflammatory cell infiltration within the lung tissues of the model group.By the 21st day,metastatic lesions manifested in the lung tissues of the model group,suggesting an approximate pre-metastatic niche maturation timeline of 14 days.Conclusion A pre-metastatic niche mouse model for Lewis lung cancer is successfully established.