摘要
探讨蛇毒金属蛋白酶抑制剂BJ46a抗肿瘤侵袭转移作用。以BJ46a为目的基因,选择杆状病毒表达系统生产重组BJ46a蛋白,ProBondTM亲和层析纯化后处理黑色素瘤细胞B16,经尾静脉接种C57BL/6小鼠,建立B16细胞实验性肺转移模型,20天后以颈椎脱臼法处死小鼠,观察肺组织病理学变化。结果表明:不同浓度重组BJ46a蛋白处理组肺部转移瘤数分别为1.1±0.83、0.9±0.7,明显低于对照组(6.3±3.00,P<0.001)和空白对照组(10.7±5.73,P<0.001),光镜下对照组肿瘤病理学征象明显,而重组BJ46a蛋白处理组瘤结节小。研究结果首次证明了重组BJ46a蛋白能抑制小鼠黑色素瘤细胞的侵袭转移,为其作为抗肿瘤侵袭转移药物的进一步研制和开发应用提供理论依据和前提条件。
Although there is emerging evidence that BJ46a can function as potent inhibitor of the SVMPs proteolytic activities, its anticancer effect on invasion and metastasis has not yet been evaluated. Inhibition effect of BJ46a on experimental pulmonary metastasis in mice inoculated with B16 melanoma ceils at the protein level was investigated. First, BJ46a was produced in baculovirus expression system. SDS-PAGE and Western blot analysis confirmed that BJ46a recombinant protein was produced by Sf9 cells infected with high-titer viral stock. Then, recombinant fusion protein was purified by ProBondTM at the point of maximal expression. B16 cells pre- treated with recombinant BJ46a injected into C57BL/6 mice via the tail lateral vein to form experimental pulmonary metastasis model . The numbers of metastatic lesions in C57BL/6 mice changed dramatically： BJ46a different concentrations of recombinant protein group were 1.1± 0.83, 0.9 ±0.7, significantly lower than the control group （6.3 ±3.00, P 〈 0.001 ） and the blank control group （ 10.7 ±5.73, P 〈 0. 001 ）. M icrostructure of the control group had obvious signs of tumor pathology, and BJ46a groups of small tumor nodules. In conclusion, the first evidence was provided that BJ46a might have a strong effect on the inhibition of the invasion and metatasis potential in vivo, and offers the theory foundation and prerequisite for development and application of anti-tumor invasion and metastasis drug.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2009年第5期28-32,共5页
China Biotechnology
基金
福建省产业技术研究开发项目(闽发改高技[2004]-686)
福建医科大学重大科技项目(XZ04002)
福建省高校创新团队培养计划(闽教科[2006]36号)资助项目
关键词
蛇毒金属蛋白酶抑制剂(BJ46a)
杆状病毒表达系统
重组蛋白
B16
侵袭转移
Snake venom metalloproteinase inhibitor （BJ46a） Recombinant protein B16 cell line Invasion and metastasis Baculovirus expression system
