因子Ⅶ-金黄色葡萄球菌肠毒素A融合蛋白的抗肿瘤效果

Anti-tumor effects of mouse factor Ⅶ-staphylococcus enterotoxin A (mfⅦ) chimeric protein in mice

目的研究鼠因子Ⅶ(mfⅦ)与金黄色葡萄球菌肠毒素A(SEA)融合蛋白的抗肿瘤生长和转移活性。方法构建表达Ⅶ因子SEA融合蛋白的腺病毒载体,用293包装细胞系制备病毒Ad/mfⅦSEA。局部荧光检测证明其表达能力和安全性后,直接应用病毒感染的293细胞以皮下注射的方式进行试验。采用小鼠205H12肺转移模型和小鼠RM1皮下肿瘤模型检测mfⅦSEA对小鼠皮下肿瘤生长和肺转移形成的抑制情况。结果腺病毒感染的293细胞诱导的二次感染只在注射局部产生,而对心、肝、肾等重要脏器无任何影响。小鼠体内实验结果显示,mfⅦSEA治疗组肺转移数(23±8)与空载体对照组(193±38)和PBS对照组(211±42)相比,差异有统计学意义(P<0.01)。在抗小鼠前列腺肿瘤的动物实验中,mfⅦSEA对皮下肿瘤的抑制非常明显,第23天时,治疗组皮下肿瘤的体积平均为(342.6±107.1)mm3,明显小于空载体对照组(2244.3±350)mm3和SEA对照组(1208.3±210)mm3。结论mfⅦSEA融合蛋白能明显抑制小鼠皮下肿瘤的生长和肺转移的形成。

Objective Mouse factor Ⅶ（mfⅦ）, ligand of tissue factor （TF）which is frequently overexpressed during neovascularlzation activated bv tumor growth, was fused to staphylococcus enterotoxin A （SEA） that mediates greater intensity of T-cell activation against tumor cells. The anti-tumor effects of the mfⅦ-SEA chimeric protein were evaluated. Methods Fusion of SEA and mfⅦ cDNA was constructed using adenovirus vector and produced in 293 packaging cell lines. The 293 cells containing the adenovirus were administered subcutaneously to mice. Fluorescence studies at the injection site and the liver were performed 3 days later. Mouse prostatic tumor RM-1 cells and mouse sarcoma MCA 205 H12 cell lines were then used in mice to create lung metastasis and subcutaneous tumor to carry out efficacy evaluation, respectively. Results Adenovirus released from the injected 293 cells only infected the subcutaneous tissue at the injection site. The in vivo experiments in mice revealed that formation of lung metastasis was strongly inhibited by the mfⅦ-SEA （23 ±8 ） compared to the vacant vector control group（ 193±38） and PBS control group（211±42） （ P 〈 0.01 ）. The mfⅦ-SEA also strongly suppressed tumor growth at the subcutaneous injection site （342.6 ±107.1 ） mm^3 compared to that of vacant vector control （2244.3±350） mm^3 and SEA （1208.3±210） mm^3 by the 23rd day. Conclusion The chimeric protein mfⅦ-SEA significantly inhibits lung metastasis formation and local tumor growth.