Visted T, Thorsen J, Thorsen F, Read T A, Ulvestad E, Engebraaten O, Sorensen D, Ylä-Herttuala S, Tyynela K, Rucklidge G, Edvardsen K, Bjerkvig R, Lund-Johansen M
Department of Anatomy and Cell Biology, University of Bergen, Bergen, Norway.
Int J Cancer. 2000 Jan 15;85(2):228-35.
The rat glioma cell lines BT4C and BT4Cn were stably transfected with the bacterial lacZ-neomycin resistance (neoR) gene construct. Both transfected (BT4ClacZ and BT4CnlacZ) and untransfected cell lines were injected intracerebrally and subcutaneously into rats. Survival time, morphology, growth rate and immunological properties of the tumors were studied. Survival time was significantly prolonged after intracerebral implantation of the transfected cell lines. No similar response was found in nude rats, indicating an immunological response towards the lacZ-neoR-transfected cells in immunocompetent animals. Morphological observations showed that the lacZ-neoR-transfected gliomas were smaller and had a distinct boundary with the normal brain tissue, whereas the parental cell lines revealed a more diffuse growth pattern. Immunostaining showed a higher proportion of immunocompetent cells infiltrating the lacZ-neoR-transfected tumors. After s.c. injection, the lacZ-neoR-transfected BT4C cell line had a prolonged lag phase before assuming a growth rate similar to that of the parental cells. The BT4CnvlacZ tumors initially grew fastest, but then disappeared within 3 weeks. A similar response was observed with mock-transfected tumor cells. A (3)HTdR-incorporation assay on spleen cells from rats transplanted s.c. with BT4CnvlacZ cells showed a 10-fold increase in cell activation as compared with rats with BT4Cn tumors. A humoral response towards the transfected cells was verified by Western-blot analyses.
大鼠胶质瘤细胞系BT4C和BT4Cn用细菌lacZ-新霉素抗性（neoR）基因构建体进行稳定转染。将转染的（BT4ClacZ和BT4CnlacZ）和未转染的细胞系脑内和皮下注射到大鼠体内。研究肿瘤的存活时间、形态、生长速率和免疫特性。转染细胞系脑内植入后存活时间显著延长。在裸鼠中未发现类似反应，表明免疫活性动物对lacZ-neoR转染细胞有免疫反应。形态学观察显示，lacZ-neoR转染的胶质瘤较小，与正常脑组织有明显边界，而亲代细胞系显示出更弥漫的生长模式。免疫染色显示有更高比例的免疫活性细胞浸润lacZ-neoR转染的肿瘤。皮下注射后，lacZ-neoR转染的BT4C细胞系在达到与亲代细胞相似的生长速率之前有一个延长的延迟期。BT4CnvlacZ肿瘤最初生长最快，但在3周内消失。用mock转染的肿瘤细胞观察到类似反应。对皮下移植BT4CnvlacZ细胞的大鼠脾细胞进行的（3）HTdR掺入试验显示，与患有BT4Cn肿瘤的大鼠相比，细胞活化增加了10倍。通过蛋白质印迹分析证实了对转染细胞的体液反应。
