论文部分内容阅读
目的 筛选能与HAb18G/CD14 7相结合的多肽 ,从中寻找该因子的肽类拮抗剂 ,抑制肝癌的恶性转移。方法 利用亲和层析法获得HAb18G/CD14 7抗原 ,在此基础上通过筛选噬菌体 12肽库得到与HAb18G/CD14 7相结合的多肽 ,以固相法合成这些多肽并用质谱仪进行鉴定。最后利用侵袭力抑制实验从中筛选拮抗肝癌细胞转移的功能多肽。结果 经纯化的HAb18G/CD14 7分子量为6 5 0 0 0 ,Westernblot验证结果阳性。用此纯化分子筛选噬菌体肽库 ,得到 9条多肽。经体外功能验证 ,9条肽抑制肝癌细胞侵袭力的活性不一 ,其中 3条有显著活性 (P <0 .0 1) ,活性最好的一条肽能使穿过Boyden小室膜的肝癌细胞数量较对照减少 90 .1%。结论 以亲和层析法和噬菌体随机肽库法相结合 ,筛选获得了有抗肝癌细胞侵袭力功能的多肽 ,此结果为抗肝癌转移肽类药物的研制奠定了基础。
OBJECTIVE: To screen for polypeptides that bind to HAb18G / CD147 and to find peptide antagonists of this factor to inhibit the malignant metastasis of hepatocellular carcinoma. Methods The HAb18G / CD147 antigen was obtained by affinity chromatography. Based on this, the peptides binding to HAb18G / CD147 were obtained by screening the phage 12 peptide library. These peptides were synthesized by solid phase method and identified by mass spectrometry. Finally, we use the invasion inhibition test to screen functional polypeptides that antagonize the metastasis of HCC cells. Results The purified HAb18G / CD14 7 had a molecular weight of 65 000 and the results of Western blotting were positive. Phage peptide library was screened with this purified molecule to obtain 9 polypeptides. After in vitro functional verification, the inhibitory activity of nine peptides on the invasiveness of hepatocellular carcinoma cells was different. Three of them had significant activity (P <0.01). The most active peptide could increase the number of hepatoma cells 90.1% less than the control. Conclusion The peptides that have the anti-invasive ability of hepatocellular carcinoma cells were screened by affinity chromatography and phage random peptide library. The results laid the foundation for the development of anti-hepatoma metastatic peptides.