邬怡然 毕振飞 黄家明 崔健 茆灿泉△.靶向FoxM1 多肽P201对人肝癌HepG2 细胞的杀伤作用及关键氨基酸分析[J].现代生物医学进展英文版,2017,17(6):1024-1028. |
靶向FoxM1 多肽P201对人肝癌HepG2 细胞的杀伤作用及关键氨基酸分析 |
Killing Effects of FoxM1 Targeted P201 Peptide against Liver Cancer HepG2Cells and Determination of Key Amino Acids in the Sequence |
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DOI: |
中文关键词: FoxM1 P201 多肽 分子对接 癌症 优化 |
英文关键词: FoxM1 P201 peptide Molecular docking Cancer Rationality |
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中文摘要: |
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;结合模体序列搜索、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60.0 滋g/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。 |
英文摘要: |
Objective:In this study, we investigated the killing effects and death pathways of FoxM1 targeted dodecapeptide P201
to human liver cancer HepG2 cells as well as key residues in the sequence.Methods:The human liver cancer HepG2 cells was used as a
model to elucidate the killing effects and death pathways of P201 peptide by MTT assay, AO-EB staining and flow cytometry. Key
residues in P201 sequence were defined by a combination of motif search, antisense peptide theory, virtual alanine mutagenesis and
molecular docking.Results:MTT assay revealed that the inhibition rate of HepG2 cells could increase to 96 % at the concentration of
60.0 滋g/mL of P201 peptide after 48 h. AO-EB staining and flow cytometry showed that early apoptosis was in a certain time- and
dose-dependent manner to P201 peptide. In addition, five key amino acids (1st, 2nd, 4th, 7th and 9th residues) were defined.Conclusion:The killing effects of P201 peptide to HepG2 cells were strong and related to apoptosis, and key residues were found in P201 sequence.
All these results provided important references and insights for the rationality of P201 and further targeted anticancer peptide drug
development. |
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