抗甲胎蛋白单链抗体与阿霉素联合对人肝癌细胞Huh7增殖的抑制作用

沈艳丽; 姬晓南; 高向东

doi:10.11665/j.issn.1000-5048.20160115

抗甲胎蛋白单链抗体与阿霉素联合对人肝癌细胞Huh7增殖的抑制作用

Combination of scFv-AFP with doxorubicin inhibits cell proliferation in hepatocellular carcinoma cell Huh7

摘要

摘要: 探讨抗人甲胎蛋白(AFP)单链抗体和阿霉素(DOX)单用或联用对人肝癌细胞Huh7细胞增殖的抑制作用。采用MTT法检测抗AFP单链抗体、阿霉素单用及联合作用对Huh7细胞增殖的影响；采用Annexin V/PI荧光双染法检测抗AFP单链抗体、阿霉素单用及联合作用对Huh7细胞凋亡的影响；采用PI单染法检测抗AFP单链抗体、阿霉素单用及联合作用对Huh7细胞周期分布的影响。结果表明，抗AFP单链抗体、阿霉素单药组与联用组均能有效抑制Huh7细胞增殖，呈剂量依赖性，且两药联用具有协同效应；抗AFP单链抗体、阿霉素单用及联用均可诱导细胞凋亡且抗AFP单链抗体(40 μg/mL)与阿霉素(0.25 μg/mL)联合组凋亡率明显高于单独给药组，具有统计学意义(P<；0.05)；抗AFP单链抗体(40 μg/mL)单独作用后，Huh7的细胞周期阻滞于G₀/G₁期，阿霉素(0.25 μg/mL)单独作用Huh7的细胞周期阻滞于S期，而当两种药物联用时，Huh7的细胞周期被阻滞于G₂/M期，抑制细胞增殖。

Abstract: This study was to investigate the inhibitory effects of single-chain variable fragment of alpha fetoprotein(scFv-AFP)in combination with doxorubicin on the proliferation of human hepatocellular carcinoma cell lines Huh7. Huh7 cells were treated with different concentrations of scFv-AFP or doxorubcin alone or their combination. The inhibitory effects were detected by MTT assay, and cycle arrest and apoptosis of Huh7 cells were analyzed by flow cytometry in different groups using PI and Annexin V/PI-staining, respectively. Results showed that scFv-AFP, doxorubicin alone or in combinations dose-dependently inhibited the proliferation of Huh7, and a synergistic effect was observed in their combined action. The combined therapy resulted in significantly higher apoptosis than those in other groups(P< 0. 05). scFv-AFP(40 μg/mL)markedly blocked the Huh7 cell progression by arresting the cells in the G₀/G₁ phase, and the percentage of cells in S phase decreased dramatically(P< 0. 05); and scFv-AFP combined with doxorubicin blocked the Huh7 cell progression by arresting the cells in G₂/M phase(P< 0. 01).

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