HGF甲基化寡核苷酸对肝癌细胞系BEL7402作用的研究

A methylated oligonucleotide inhibits HGF expression in liver cancer cell line BEL-7402

目的 :肝细胞生长因子 (HGF)能促进肿瘤生长 ,转移及血管形成 ,利用甲基化灭活基因的原理 ,探讨HGF启动子甲基化寡核苷酸对肝癌细胞系BEL 74 0 2的作用。方法 :分别用甲基化寡核苷酸、非甲基化寡核苷酸、PBS处理BEL 74 0 2细胞 ,MTT检测肿瘤抑制率 ,RT PCR检测HGFmRNA表达状况 ,流式细胞仪观察细胞凋亡率。结果 :当甲基化寡核苷酸 4 μM、6 μM、8μM、10 μM时 ,肿瘤抑制率分别为4 7%、5 0 %、6 4 %、75 %。RT PCR显示其最早在 8h发挥作用 ,维持 72h。甲基化寡核苷酸处理组的细胞凋亡率显著高于其它各组 (χ2 =94 ,P <0 .0 1)。结论 :HGF甲基化寡核苷酸能够抑制肝癌细胞BEL 74 0

Objective: The tumour growth , metastasis, vascularization can be promoted by HGF,the gene can be inactivated through its methylation.This experiment was carried out to explore whether the methylated oligonucleotide of HGF promoter had any effect on liver cancer cell line BEL-7402.Methods:The BEL-7402 cells were treated by the methylated oligonucleotide, unmethylated oligonucleotide or PBS solutions. The tumour suppression rate was measured by MTT, the expression of HGFmRNA was detected by RT-PCR,the apoptosis rate was detected by flow cytometer.Results: When the concentrations of the methylated oligonucleotide was 4μM,6μM,8μM,10μM,the tumor suppressionrate were47%,50%,64%,75%respectively.The result of RT-PCR showed that the methylated oligonucleotide had anti-tumour effect as early as 8 hours and last for 72 hours. The apoptosis rate observed by flow cytometer was high in the methylated oligonucleotide treated goups than that of any other groups.Conclusion:The methylated oligonucleotide of HGF promoter has anti-tumour effect on live cancer cell line BEL-7402.