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目的:研究从日本蛇菰中分离的天然多酚化合物1,2,6-三-O-没食子酰基-β-D-吡喃葡萄糖(BJA32531)对人肝癌细胞HepG2增殖的抑制作用以及对miRNA表达的影响。方法:采用CCK-8的方法检测HepG2细胞的增殖;用流式细胞法检测HepG2细胞的凋亡;采用miRNA芯片分析方法测定HepG2细胞的miRNA表达以及用RT-PCR的方法验证miRNA的表达。结果:BJA32531以时间和剂量依赖的方式抑制HepG2细胞的增殖;流式细胞结果表明:该化合物能引起HepG2细胞的凋亡。miRNA芯片结果显示,BJA32531能诱导HepG2细胞19个miRNA的表达上调以及85个miRNA的表达下调。RT-PCR验证了化合物诱导的let-7a和miR-10b的上调以及miR-132和miR-125b的下调与芯片的结果一致。结论:BJA32531抗HepG2细胞增殖作用的机理可能与调控miRNA的表达有关。
Aims: To study the inhibitory effect of natural polyphenol compound 1,2,6-tri-O-galloyl-β-D-glucopyranose (BJA32531) isolated from Japanese snakehead fish on the proliferation of HepG2 human hepatocellular carcinoma cells and its effect on miRNA expression Impact. Methods: The proliferation of HepG2 cells was detected by CCK-8. The apoptosis of HepG2 cells was detected by flow cytometry. The miRNA expression of HepG2 cells was determined by miRNA microarray analysis and the miRNA expression was verified by RT-PCR. Results: BJA32531 inhibited the proliferation of HepG2 cells in a time and dose-dependent manner. The results of flow cytometry showed that the compound could induce the apoptosis of HepG2 cells. Results of miRNA microarray showed that BJA32531 induced up-regulation of 19 miRNAs and down-regulation of 85 miRNAs in HepG2 cells. RT-PCR validated compound-induced up-regulation of let-7a and miR-10b as well as down-regulation of miR-132 and miR-125b in agreement with chip results. Conclusion: The mechanism of BJA32531 against HepG2 cell proliferation may be related to the regulation of miRNA expression.