全反式维甲酸诱导HepG2细胞分化和降低软琼脂克隆形成

Differentiation induced and colony formation breakdown in HepG2 cells treated with all-trans-retinoic acid

目的研究全反式维甲酸(ATRA)体外对人肝癌细胞株(HepG2)分化、软琼脂克隆形成的影响及其机制。方法ATRA处理HepG2,光镜下观察细胞形态,并检测γ-谷氨酰转肽酶(γ-GT)、甲胎蛋白(AFP)等分化指标的变化;四甲基偶氮唑盐(MTT)法分析ATRA对HepG2增殖的影响;软琼脂培养法观察HepG2的克隆形成能力;Western blot检测骨桥蛋白(OPN)表达变化。结果ATRA显著抑制肝癌细胞增殖并诱导细胞分化,使代表肝细胞恶变的γ-GT比活力、AFP分泌量明显下降。ATRA可降低HepG2软琼脂克隆形成能力并使OPN表达减少。结论ATRA是HepG2有效的分化诱导剂,并可抑制HepG2增殖,其分子机制可能与OPN表达减少有关,为ATRA诱导分化治疗肝癌提供了实验依据。

Objective To study the differentiation and the ability of colony formation of human hepatoma HepG2 cells induced by all-trans-retinoic acid （ATRA） and its mechanism. Methods HepG2 was treated with different concentrations of ATRA. The morphology of HepG2 was observed by light microscope and the diversity of γ-glutamyl transpeptidase（γ-GT） and alpha-fetoprotein（AFP） was detected by kinetics analysis. The cell proliferation of HepG2 was analyzed by MTT assay. The ability of colony formation was assaged by soft agar. The expression of osteopontin （OPN） was studied by Western blot. Results The proliferation of HepG2 cells, the specific activities of γ-GT and the secretary amount of AFP significantly decreased;The ability of colony formation significantly broken down and the Western blot indicated a significant down-regulation of OPN in HepG2 treated with ATRA. Conclusion ATRA is effective to induce differentiation and can inhibit proliferation of HepG2 by down-regulation of OPN.