摘要
目的探讨聚腺苷酸二磷酸核糖转移酶-1（PARP-1）抑制剂4-氨基-1,8-萘二胺（4-AN）对三氧化二砷（ATO）治疗肝癌的敏感性影响及其相关机制。方法培养人肝癌细胞HepG2,分为单独ATO用药组和联合用药组（4-AN＋ATO）,采用MTT实验、群体倍增实验和克隆形成实验比较两组细胞存活、增殖情况;应用ELISA、单细胞凝胶电泳（彗星实验）及微核实验分别检测细胞内8-羟基鸟嘌呤（8-OH-dG）含量,DNA链断裂、修复以及染色体损伤情况。结果当ATO浓度为2~10μmol/L时,联合用药组细胞存活率和集落形成率低于ATO组（P〈0.05）,ATO 2~50μmol/L时,联合用药组细胞群体倍增时间高于ATO组（P〈0.05）;在ATO 2~20μmol/L浓度范围内,彗星细胞尾长（L-Tail）、尾距（OTM）和8-OH-dG含量较ATO组高,差异均有统计学意义（P均〈0.05）;彗星实验检测损伤修复结果显示,联合用药组细胞的DNA损伤修复效率低于ATO组（P〈0.05）;微核实验结果显示当ATO浓度为5~20μmol/L时,联合用药组微核细胞率高于ATO组（P〈0.05）。结论 PARP-1抑制剂4-AN能显著增强ATO杀伤肝癌细胞的敏感性,其机制与抑制肝癌细胞DNA损伤修复有关。
Objective To study the effect and mechanism of 4-amino-1,8-naphthalimide（4-AN）on the sensitive effect of arsenic trioxide（ATO）in hepatocellular carcinoma cells.Methods Hepatocellular carcinoma HepG2 cells were divided into two groups according to whether they were treated with 4-AN or not.Cell viability was evaluated by MTT assay,population doubling experiment and colony formation assay;genic mechanism was explored by 8-OH-dG assay,single cell gel electrophoresis（comet assay）and micronucleus test.Results At 2-10μmol/L concentration of ATO,the cell viability and colony formation efficiency of the combination group（4-AN＋ATO）were significantly lower than that of the ATO group（P〈0.05）;moreover,the tail-length（L-Tail）and olive tail moment（OTM）in comet assay were notablely higher than that of the ATO group（P〈0.05）.At 2-20μmol/L concentration of ATO,the population doubling time and 8-OH-dG in combination group were significantly higher than that of ATO group（P〈0.05）.Results from DNA damage repair assay showed that the efficiency of DNA damage repair in combination group was remarkably lower than that of ATO group（P〈0.05）.At 5-20μmol/L concentration of ATO,the frequency of micronucleated cells in combination group was significantly higher than that of ATO group（P〈0.05）.Conclusion 4-AN can significantly increase the sensitivity of ATO in treatment with hepatocellular carcinoma cells and prevent DNA damage repair may be a primary mechanism for this effect.
出处
《四川大学学报（医学版）》
CAS
CSCD
北大核心
2015年第2期163-168,共6页
Journal of Sichuan University(Medical Sciences)
基金
国家自然科学基金面上项目(No.81172632)资助
