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目的:初步探究靶向FoxM1多肽P201对人肝癌细胞的杀伤作用、死亡途径以及寻找多肽序列中关键的氨基酸残基。方法:本研究选取人肝癌HepG2细胞为主要研究对象,采用MTT法、AO-EB双染和流式细胞术检测P201多肽对HepG2细胞的杀伤作用和死亡途径,结合模体序列捜索、反义肽氨基酸、虚拟丙氨酸突变和分子对接等方法:确定P201多肽的关键氨基酸。结果:MTT法检测60.0μg/mL P201多肽对HepG2细胞作用48 h抑制率高达96%,形态观察和定量测定显示细胞早期凋亡的发生与P201多肽作用时间和剂量呈一定依赖关系;共确定5个氨基酸(第1、2、4、7、9位残基)为P201多肽的关键氨基酸残基。结论:初步揭示P201多肽对HepG2细胞的强杀伤作用与细胞凋亡相关并确定关键氨基酸为多肽分子的优化和多肽抗癌靶向药物的进一步研发提供了重要的依据与参考。
OBJECTIVE: To investigate the killing effect of Foxp1 targeting P201 on human hepatoma cells, the pathways of death and the search for the key amino acid residues in the polypeptide sequence. Methods: Human HepG2 cells were selected as the main research object. The killing effect and death pathway of P201 peptide on HepG2 cells were detected by MTT assay, AO-EB double staining and flow cytometry. Combined with motif sequence and antisense peptide Amino acids, virtual alanine mutations and docking methods such as molecular docking: P201 polypeptide to determine the key amino acids. Results: The inhibitory rate of 60.0 μg / mL P201 peptide on HepG2 cells after 48 h treatment was 96%. MTT assay showed that the early apoptosis of cells was dependent on the time and dosage of P201 polypeptide. A total of 5 Amino acids (residues 1, 2, 4, 7 and 9) are the key amino acid residues of the P201 polypeptide. CONCLUSION: The preliminary study reveals that P201 peptide has a strong anti-cancer effect on HepG2 cells, which is related to apoptosis and it is important to determine the optimization of key amino acids and further research and development of peptide anticancer drugs.