【摘要】 目的探讨3株不同来源的铜绿假单胞菌分泌性蛋白的抗肿瘤活性。方法将3株不同来源的铜绿假单胞菌经LB培养基对其过夜静置培养,离心获得上清液,采用硫酸铵盐析沉淀蛋白质,再经PBS透析除盐。然后将蛋白作用于人肝癌细胞Hep-G2、宫颈癌细胞HeLa、肺癌细胞A549和人永生化表皮细胞HaCaT,CCK-8检测其对细胞的毒性作用,吉姆萨染色观察凋亡细胞形态变化。结果 SDS-PAGE证实成功获得不同分子量的分泌蛋白,CCK-8结果显示混合蛋白对3种不同肿瘤细胞的生长均有不同程度的抑制作用,且其抑制作用存在一定的时间和浓度依赖性,但对人正常细胞无明显抑制作用。吉姆萨染色初步显示细胞形态为凋亡状态。结论初步证实该3株铜绿假单胞菌产生的分泌性蛋白具有不同程度的抗肿瘤作用,为进一步分离纯化具有抗肿瘤活性的单一蛋白质及研究其抗肿瘤机制提供依据。更多还原

【Abstract】 Objective To explore the antitumor activity of secretory proteins produced by three strains of Pseudomonas aeruginosaisolates from different sources.Methods LB medium was used to culture the three strains overnight without shaking.The culture liquid was centrifuged,and the proteins were extracted from the supernatant through ammonium sulfate precipitation and PBS dialysis.Three human tumor cell lines and one normal cell line as well as CCK-8 were used to detect the cytotoxicity of the proteins.Results The acquisition of secretory proteins with different molecular weights was confirmed by SDS-PAGE.The CCK-8 results showed that they had inhibitory effects on the growth of tumor cells,while no evident cytotoxicity on normal cells.Giemsa dyeing showed cells in apoptosis.Conclusion The secretory proteins produced by these three P.aeruginosahave cytotoxic effects on human tumor cell lines.Based on this,we can try to isolate single proteins with antitumor activity and study their antitumor mechanisms.更多还原