小细胞肺癌原代细胞脑转移模型研究初探

Establishment of primary cell brain metastasis model of small cell lung cancer:a preliminary study

目的本研究拟分离培养保留原始肿瘤的重要特征和功能的小细胞肺癌(small cell lung cancer,SCLC)原代细胞,并在此基础上构建SCLC原代细胞脑转移小鼠模型,为SCLC脑转移患者的药物研发和评估提供更贴近临床的动物模型。方法收集临床病人的新鲜SCLC肿瘤组织样本进行小鼠皮下移植,通过小鼠的荷瘤情况、肿瘤组织大小和肿瘤生长曲线的结果,确定小细胞肺癌人源肿瘤异种移植(patient-derived xenograft,PDX)模型的构建情况。并对成瘤后的肿瘤组织进行原代细胞分离培养,通过细胞划痕实验和增殖实验检测细胞的增殖和侵袭能力。通过颈动脉注射原代细胞构建SCLC脑转移模型,并通过临床表现和病理观察确定转移模型是否成功。结果(1)成功构建SCLC皮下PDX模型,肿瘤传代时间约为56 d,肿瘤生长曲线呈对数生长趋势。(2)成功分离筛选出具备成瘤性的SCLC原代L0细胞,并证实该L0细胞具备较强的细胞迁移和增殖能力。(3)颈动脉注射法成功构建SCLC脑转移模型,并分离出具备脑转移特性的SCLC脑转移B0细胞。结论本研究利用分离培养SCLC原代细胞,成功构建SCLC原代细胞的脑转移的模型。

Objective To isolate and culture primary small cell lung cancer(SCLC)cells that retain the important characteristics and functions of the original tumor,and use them to construct a brain metastasis model of SCLC,as an animal model for drug development and the evaluation of patients with SCLC brain metastasis.Methods Fresh SCLC tumor tissue samples were collected from patients and transplanted subcutaneously into mice.The result ing patient-derived xenograft(PDX)mouse model of SCLC was assessed in terms of tumor presence,tumor size,and tumor growth.Primary cells were isolated and cultured from the tumor tissue after tumor formation,and their proliferation and invasion abilities were detected by cell scratch and proliferation tests.An SCLC brain metastasis model was constructed by injecting primary cells into the carotid artery,and the model was assessed in terms of its clinical manifestations and pathological observations.Results(1)A subcutaneous PDX model of SCLC was constructed successfully.The tumor passage time was about 56 days,and the tumor growth curve showed a logarithmic growth trend.(2)Primary L0 SCLC cells with tumorigenicity were successfully isolated and screened,and demonstrated strong cell migration and proliferation abilities.(3)An SCLC brain metastasis model was successfully constructed by carotid injection,and SCLC B0 cells with brain metastasis characteristics were isolated.Conclusions Primary SCLC cells were successfully isolated and cultured from SCLC tumor tissue samples,and a brain metastasis model using primary SCLC cells was successfully constructed.