摘要
目的观察多株肝癌细胞中TIMP-3 CpG岛的甲基化状况,观察甲基化药物5-杂氮-2′脱氧胞苷对肝癌细胞株中TIMP-3基因表达及其CpG岛甲基化程度的影响。方法运用MSP定性检测8株肝癌细胞中TIMP-3 CpG岛甲基化状况。使用去甲基化药物5-杂氮-2′脱氧胞苷对甲基化阳性的细胞株进行干预,观察干预组与对照组TIMP-3基因表达差异,并使用焦磷酸测序技术定量检测CpG岛甲基化差异。结果在8株肝癌细胞系中C3A、Hep-3B、HepG2 3株检测到TIMP-3 CpG岛甲基化阳性。经去甲基化药物干预后,在这3株肝癌细胞中均发现TIMP-3 mRNA的表达较对照组有明显上调(P<0.05),CpG岛的甲基化率下降(P<0.05)。结论甲基转移酶抑制剂能显著降低肝癌细胞株TIMP-3 CpG岛甲基化程度及上调TIMP-3 mRNA的表达。
Objective The aim of this study is to investigate the CpG island methylation status of TIMP- 3 in HCC cell lines, to treate the HCC cells that exhibit positive methylation with DNA methyltransferase in- hibitor （5-Aza-CdR）, and to observe the influence of 5-Aza-CdR on the expression of TIMP-3 mRNA and its CpG island methylation degree. Methods TIMP-3 CpG methylation status of HCC cells were examined using MSP assay. Methylation-positive HCC cells were treated with DNA methyltransferase inhibitor （5-Aza- CdR） （intervention group）, and the difference of TIMP-3 mRNA expression was observed between interven- tion group and control group. Pyrosequencing method was applied to quantitatively examine the CpG island methylation percentage. Results The methylation of TIMP-3 CpG islands in C3A, HepG-2, and Hep-3B was positive. The results showed that TIMP-3 mRNA expression was increased （P 〈 0.05） and the methylation level was decreased （P 〈 0.05） in the intervention group compared to control group. Conclusion 5-Aza-CdR could remarkably reduce the level of the CpG island methylation and increase the expression of TIMP-3 mR- NA in HCC cells.
出处
《现代消化及介入诊疗》
2013年第6期340-343,共4页
Modern Interventional Diagnosis and Treatment in Gastroenterology
基金
教育部博士学科点专项基金(201144231100 05)
广东省科技计划项目(2010B031600308)
