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目的:探讨人血清淀粉样蛋白A家族成员在肿瘤转移中的作用和机制。方法:表达和纯化人SAA1-GST、SAA2-GST、SAA3-GST和SAA4-GST融合蛋白,采用体外迁移实验,观察和比较人SAA1-GST、SAA2-GST、SAA3-GST、SAA4-GST和GST对lewis肺癌(lewis lung carcinoma LLC)细胞、Ba/mTLR4/mMD2细胞及其对照Ba/κB细胞的迁移作用。通过蛋白质印迹法观察人SAA1-GST、SAA2-GST、SAA3-GST、SAA4-GST和GST作用后LLC细胞的IkB和pIkB蛋白表达。结果:成功表达和纯化人SAA1-GST、SAA2-GST、SAA3-GST和SAA4-GST融合蛋白。与GST对照组相比,人的4种融合蛋白均能显著促进LLC细胞的迁移,P<0.05;人SAA1-GST、SAA2-GST、SAA3-GST和SAA4-GST融合蛋白与GST相比,均能明显的促进Ba/mTLR4/mMD2细胞迁移(P<0.05),而对对照组Ba/κB细胞无此作用,P>0.05。蛋白质印迹法结果显示,人SAA1-GST、SAA2-GST、SAA3-GST和SAA4-GST较GST作用后LLC细胞pIKB蛋白表达明显增强,P<0.05。结论:人SAA家族成员都能促进肿瘤细胞的迁移,并且可能是通过激活由TLR4(toll like receptor 4)介导的NF-κB信号转导通路发挥该作用。
Objective: To investigate the role and mechanism of human serum amyloid A family members in tumor metastasis. METHODS: SAA1-GST, SAA2-GST, SAA3-GST and SAA4-GST fusion protein were expressed and purified in vitro. The migration and expression of SAA1-GST, SAA2- Migration of lewis lung carcinoma LLC cells, Ba / mTLR4 / mMD2 cells and their control Ba / κB cells. The expression of IkB and pIkB proteins in LLC cells after human SAA1-GST, SAA2-GST, SAA3-GST, SAA4-GST and GST treatment were observed by Western blotting. Results: The human SAA1-GST, SAA2-GST, SAA3-GST and SAA4-GST fusion proteins were successfully expressed and purified. Compared with GST control group, all four fusion proteins of human could significantly promote the migration of LLC cells, P <0.05; Compared with GST, the fusion protein of SAA1-GST, SAA2-GST, SAA3-GST and SAA4- Can significantly promote the migration of Ba / mTLR4 / mMD2 cells (P <0.05), but no effect on Ba / κB cells in control group (P> 0.05). The result of Western blot showed that the expression of pIKB protein in LLC cells was significantly increased after human SAA1-GST, SAA2-GST, SAA3-GST and SAA4-GST compared with GST, P <0.05. Conclusion: Human SAA family members can promote the migration of tumor cells, and may play its role through activation of NF-κB signaling pathway mediated by TLR4 (toll like receptor 4).