重组BJ46a 蛋白抑制B16 细胞的体外侵袭及转移

目的 探讨蛇毒金属蛋白酶抑制剂BJ46a的抗肿瘤侵袭及转移作用. 方法 以蛇毒金属蛋白酶抑制剂BJ46a为目的基因,选择杆状病毒表达系统生产重组BJ46a蛋白,纯化并测定其生物学活性;利用Alamar blue法及Transwell小室分析法评价重组蛋白对小鼠黑色素瘤细胞B16体外生长、黏附、运动和侵袭能力的影响(n＝3). 结果 经ProBond亲和层析纯化出的重组BJ46a蛋白具有抑制基质金属蛋白酶(MMPs)的活性;经重组BJ46a蛋白处理的B16 细胞穿过 Matrigel的细胞数明显低于对照组(P<0.01),抑制率为84.8%. 结论重组BJ46a蛋白能抑制B16细胞的侵袭转移.

Recombinant BJ46a protein exerts inhibition effect on invasion and metatasis of B16 cells in vitro

P>Objective To investigate the effects of BJ46a on invasion and metastasis of melanoma cells. Methods A snake venom metalloproteinase inhibitor BJ46a was produced in baculovirus expression system. Then, recombinant fusion protein was purified by ProBond at the point of maximal expression, SDS-PAGE and Western blotting were used to detect the protein expression. The inhibition activity of matrix metalloproteinasel(MMPs) was detected by purified BJ46a protein. The abilities of tumor cell proliferation and adhension were determined by Alamar blue assay(EM>n/EM>=3).The ability of tumor cell invasion was identified by transwell chamber EM>in vitro/EM>(EM>n/EM>=3). Results The activity of MMPs could be reduced by purified BJ46a protein. The ability of B16 cells treated with recombinant BJ46a to invade the reconstituted basement membrane decreased significantly(EM>P/EM><0.01)，inhibition ratio was 84.8%, but the proliferation and adhension abilities had no change.Conclusion BJ46a might has a strong inhibition effect on the invasion and metatasis potential of B16 cells EM>in vitro/EM>./P>