小鼠肺原位移植瘤及淋巴结转移模型的建立

Establishment of a mouse model of orthotopic lung cancer and lymph node metastasis

目的建立一种小鼠肺原位移植瘤及淋巴结转移模型,模拟肺癌发生发展过程中淋巴结转移,进行肺癌转移前微环境中免疫细胞的研究。方法培养小鼠Lewis肺癌细胞,经胸壁细胞注射法构建小鼠肺原位移植瘤模型,分别于肿瘤移植后第7、14、21和28天取材,苏木素-伊红(HE)染色和免疫组化法(immunocytochemistry,IHC)观察原位移植瘤、淋巴结组织结构及B细胞变化。结果HE染色观察到随着肺癌的发展,淋巴结内生发中心、淋巴管及血管增多,发生肿瘤转移的淋巴结增大变形并有肿瘤细胞浸润。IHC结果同样表明原位肺癌模型小鼠淋巴结B细胞密度增加。结论成功构建C57BL/6J小鼠原位肺癌模型,该模型的建立为今后肺癌淋巴结转移前微环境中免疫细胞的相关研究奠定了基础。

Objective Establishment of a mouse model of orthotopic lung cancer and lymph node metastasis to simulate lymph node metastasis during the occurrence and development of lung cancer and to study the immune cells in the microenvironment before lung cancer metastasis occurs.Methods Mouse Lewis lung cancer cells were cultured and then injected through the chest walls of mice to establish an orthotopic mouse model of lung cancer.Samples were taken at 7,14,21,and 28 days after tumor transplantation,and the changes in the orthotopically transplanted tumors,lymph node tissue structure,and B cells were observed by HE staining and immunohistochemistry.Results The orthotopic C57BL/6J mouse model of lung cancer was successfully constructed,and the mediastinal lymph nodes were collected.The changes in the germinal center,lymphatic vessels,and blood vessels within the lymph nodes as well as structural alterations of lymph nodes associated with tumor metastasis and infiltration of tumor cells were observed during the progression of lung cancer using HE staining.Immunohistochemistry showed an increase in the density of B cells in the lymph nodes of the model group.Conclusions The establishment of this model lays a foundation for the future study of immune cells in the microenvironment of lung cancer before lymph node metastasis occurs.