Selective cytotoxicity of goniothalamin against hepatoblastoma HepG2 cells.
Selective cytotoxicity of goniothalamin against hepatoblastoma HepG2 cells.
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DOI：
10.3390/molecules16042944
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发表时间：
2011-04-06
期刊：
影响因子：
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通讯作者：
Alitheen NB
中科院分区：
文献类型：
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作者：
Al-Qubaisi M;Rozita R;Yeap SK;Omar AR;Ali AM;Alitheen NB
肝癌已成为高死亡率的主要癌症类型之一，而目前化疗中使用的细胞毒药物对肝癌的治疗效果不佳。本研究的目的是检测眼镜蛇甲素对人肝母细胞瘤HepG2细胞和正常肝Chang细胞的体外细胞毒作用。采用四甲基偶氮唑盐比色法、乳酸脱氢酶漏出法、细胞周期流式细胞仪PI分析、BrdU增殖酶联免疫吸附试验和台盼蓝拒染试验检测了该药对人肝癌细胞株HepG2和肝常细胞的细胞毒作用。Goniothalamin对HepG2细胞有选择性的抑制作用[MTT法IC50=4.6(±0.23)µM；LDH法72 h IC50=5.20(±0.01)µM]，对Chang细胞的敏感性较差[MTT法IC50=35.0(±0.09)µM；LDH法72 h IC50=32.5(±0.04)µM]。台盼蓝拒染实验显示，72小时后，细胞活力指数分别为52±1.73%和62±4.36%。从BrdU掺入的减少可以看出，GNP的细胞毒性与其对DNA合成的抑制有关。72h时，最低浓度(2.3ug/L)的L对正常肝Chang细胞的增殖抑制率为97.6%，而对HepG2细胞的增殖抑制率为19.8%。此外，流式细胞仪分析细胞周期显示，当归甲素可引起亚二倍体细胞凋亡聚集和不同程度的G2/M期阻滞。Goniothalamin通过抑制细胞增殖和诱导细胞凋亡而选择性地杀伤肝癌细胞。这些结果表明，对肝母细胞瘤HepG2细胞具有潜在的细胞毒作用。
Liver cancer has become one of the major types of cancer with high mortality and liver cancer is not responsive to the current cytotoxic agents used in chemotherapy. The purpose of this study was to examine the in vitro cytotoxicity of goniothalamin on human hepatoblastoma HepG2 cells and normal liver Chang cells. The cytotoxicity of goniothalamin against HepG2 and liver Chang cell was tested using MTT cell viability assay, LDH leakage assay, cell cycle flow cytometry PI analysis, BrdU proliferation ELISA assay and trypan blue dye exclusion assay. Goniothalamin selectively inhibited HepG2 cells [IC50 = 4.6 (±0.23) µM in the MTT assay; IC50 = 5.20 (±0.01) µM for LDH assay at 72 hours], with less sensitivity in Chang cells [IC50 = 35.0 (±0.09) µM for MTT assay; IC50 = 32.5 (±0.04) µM for LDH assay at 72 hours]. In the trypan blue dye exclusion assay, the Viability Indexes were 52 ± 1.73% for HepG2 cells and 62 ± 4.36% for Chang cells at IC50 after 72 hours. Cytotoxicity of goniothalamin was related to inhibition of DNA synthesis, as revealed by the reduction of BrdU incorporation. At 72 hours, the lowest concentration of goniothalamin (2.3 µL) retained 97.6% of normal liver Chang cells proliferation while it reduced HepG2 cell proliferation to 19.8% as compared to control. Besides, goniothalamin caused accumulation of hypodiploid apoptosis and different degree of G2/M arrested as shown in cell cycle analysis by flow cytometry. Goniothalamin selectively killed liver cancer cell through suppression of proliferation and induction of apoptosis. These results suggest that goniothalamin shows potential cytotoxicity against hepatoblastoma HepG2 cells.