丹参注射液对人肝癌细胞14-3-3σ基因甲基化的影响

Effect of Radix Salviae Miltiorrhizae on DNA Methylation of 14-3-3σ in Human Hepatocellular Carcinoma

目的观察肝癌组织14-3-3σ基因表达及其启动子区域甲基化程度的改变,丹参注射液对人肝癌细胞株Huh-7细胞14-3-3σ基因表达及其启动子区域甲基化的影响。方法收集2014—1月至2014年12月36例肝癌患者的肝癌组织及其癌旁组织,分别分离其RNA及DNA,针对14-3-3σ基因表达及其启动子区域甲基化程度进行定量分析;使用丹参处理人肝癌细胞株Huh-7细胞,观察丹参注射液处理后人肝癌细胞株Huh-7细胞侵袭能力及其14-3-3σ基因表达及其启动子区域甲基化程度的改变。结果相较于癌旁组织,肝癌组织14-3-3σ基因的表达量显著降低,且基因组DNA启动子区域多个CpG位点呈连续性的甲基化水平增高;丹参注射液处理后人肝癌细胞株Huh-7细胞侵袭能力显著降低;伴随着14-3-3σ基因表达量增加,且其启动子区域的多个CpG位点呈连续性的甲基化水平降低。结论肝癌组织14-3-3σ基因低表达与其启动子区域的高甲基化水平相关;丹参注射液能抑制通过抑制14-3-3σ基因启动子区域的多个CpG位点的甲基化,促进抑癌基因14-3-3σ表达,从而降低人肝癌细胞株Huh-7细胞的侵袭能力。

Objective To investigate the expression of 14-3-3σ and the methylation of its promoter in hepato-cellular carcinomas （HCC）. To observe the effect that Salvia Miltiorrhiza on 14-3-3σ. Methods Hepatic cancer tissue and normal liver tissue were collected from 36 HCC patients between January 2014 and December 2014 to quantify the expression and methylation of 14-3-3σ. HCC cell line Huh-7 was treated with radix salvia miltior-rhiza to observe the change of the gene expression and methylation of 14-3-3σ. Results The expression of 14-3-3σ significantly decreased in cancer tissue compared to that in the paired normal liver tissue. The DNA promoter CpG methylation was also increased. After treatment of radix salvia miltiorrhiza, the invasion of Huh-7 cells was reduced; the expression of 14-3-3σ was increased and the methylation was decreased. Conclusion The low ex-pression of 14-3-3σ is correlated with a high methylation level in HCC. Radix salvia miltiorrhiza can suppress the invasion of Huh-7 cells via inhibiting the promoter CpG methylation and increasing the expression of cancer sup-pressor gene 14-3-3σ.