Nakashima E, Kubota Y, Matsushita R, Ozaki E, Ichimura F, Kawahara S, Nakanishi I, Kuno K, Matsushima K
Hospital Pharmacy, Kanazawa University, Japan.
Pharm Res. 1998 May;15(5):685-9. doi: 10.1023/a:1011906600304.
In order to evaluate the possibility of synergistic antitumor gene therapy by the gene delivery of monocyte chemotactant protein-1 (MCP-1/MCAF/IE), the effect of a biological response modulater for macrophages on tumor progression of gene transfected tumor cells was studied.
Cachexia-inducing adenocarcinoma cells (cell line colon 26, clone 20) were transfected with either a control plasmid or MCP-1 cDNA.
The production of MCP-1 reached 70-80 ng/ml in vitro when transfectant cells were cultured at a cell density of 1 x 10(5) cells/ml for 3 days. Transfection of MCP-1 cDNA did not affect the growth rate in vitro. Also, MCP-1-transfectants formed tumors after intra-footpad inoculation similar in size to the parental cells. The number of infiltrating macrophages in the primary tumor of the transfectant rapidly increased from the 3rd to 5th day after inoculation as revealed by immunohistochemical staining using an antibody against mouse macrophages. An earlier, greater, but no longer-lasting increase in tumor-infiltrating macrophages was induced in tumors by MCP-1 transfection was compared to that induced by the parent cells. On the 10th day after the inoculation, the tumor-infiltrating macrophages in mice inoculated MCP-1 transfectants were decreased to a level similar to that of the parent cells. Groups of mice were treated intraperitoneally with LPS at different times after the inoculation. Tumor cells producing high levels of MCP-1 were significantly lysed by macrophages treated with LPS, whereas parental or control transfected cells were not. Conclusions. Combination immunotherapy can provide a rationale for the application of MCP-1 treatment to increase immunological responses to cancer.
为了评估通过单核细胞趋化蛋白-1（MCP-1/MCAF/IE）基因递送进行协同抗肿瘤基因治疗的可能性，研究了一种巨噬细胞生物反应调节剂对基因转染肿瘤细胞肿瘤进展的影响。
将诱导恶病质的腺癌细胞（结肠26细胞系，克隆20）用对照质粒或MCP-1 cDNA进行转染。
当转染细胞以1×10⁵个细胞/毫升的细胞密度培养3天时，体外MCP-1的产生量达到70 - 80纳克/毫升。MCP-1 cDNA的转染不影响体外生长速率。此外，MCP-1转染细胞经足垫内接种后形成的肿瘤大小与亲代细胞相似。使用抗小鼠巨噬细胞抗体进行免疫组织化学染色显示，接种后第3天至第5天，转染细胞原发性肿瘤中浸润的巨噬细胞数量迅速增加。与亲代细胞诱导的情况相比，MCP-1转染诱导肿瘤浸润巨噬细胞的增加更早、程度更大，但持续时间不长。接种后第10天，接种MCP-1转染细胞的小鼠体内肿瘤浸润巨噬细胞数量降至与亲代细胞相似的水平。接种后不同时间给小鼠腹腔注射LPS。产生高水平MCP-1的肿瘤细胞被LPS处理的巨噬细胞显著裂解，而亲代或对照转染细胞则未被裂解。结论。联合免疫疗法可为应用MCP-1治疗以增强对癌症的免疫反应提供理论依据。
