| 摘要: |
| 目的 检测胃癌细胞株MGC803内乳铁蛋白（LTF）基因的甲基化和表达情况，探讨LTF的DNA 启动子区甲基化异常
与其在胃癌内表达的相关性。方法 用亚硫酸氢盐测序PCR 方法检测MGC803 启动子区甲基化状态，使用real-time PCR 和Western blot 分别检测LTF 在MGC803内的mRNA 和蛋白表达水平。同时使用去甲基化剂5-aza-CdR 处理胃癌细胞株，观察给药前后的DNA 甲基化状态和mRNA 表达情况。结果 MGC803 启动子甲基化率达59.8%，使用5-aza-CdR 处理的两组（2滋mol/L组和10滋mol/L组）和空白对照组甲基化率差异、LTF 的mRNA表达差异均有统计学意义（均P＜0.05），并且随着MGC803启动子甲基化程度的下降，其mRNA 和蛋白表达增加；而不同浓度药物处理组间差异均无统计学意义（均P＞0.05）。结论 LTF 基因在胃癌细胞株MGC803 内表现为较高甲基化状态，而且LTF的表达和其启动子区甲基化情况相关，使用去甲基化剂能逆转DNA 的甲基化情况从而使其mRNA 重新表达。 |
| 关键词: LTF 基因 胃肿瘤 5-杂氮脱氧胞苷 DNA 甲基化 CpG岛 |
| DOI： |
| 分类号: |
| 基金项目:温州市科技局项目 |
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| Expression of LTF gene and its promotor region methylation in human gastric cancer cell lines MGC803 |
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CHEN Xuyan,ZHUGE Xiaoju,CHEN Renpin,HUANG Zhimin
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the First Affiliated Hospital of Wenzhou Medical University
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| Abstract: |
| Objective To investigate the expression of LTF (lactotransferrin) gene and its promoter region in human gastric cancer cell line MGC803. Methods The expression of LTF mRNA and protein levels in MGC803 cells were detected by real-time PCR and Western blot, respectively; LTF promotor methylation status was determined by Bisulfite genomic sequencing PCR (BSP). MGC803 cells were treated with 5-aza-CdR to reverse methylation status, Results LTF promotor methylation rate in MGC803 was 59.8%. Compared to control group, methylation status and mRNA expression of LTF in 5-aza-CdR treatment groups(2滋mol/L and 10滋mol/L) were reversed(P＜0.05); however there was no difference between different treatment groups(P＞0.05). Conclusion The LTF methylation in promoter region is high and LTFmRNA expression in MGC803 cells is low, 5-aza-CdR can reverse the methylation status and up-regulate LTF mRNA expression. |
| Key words: Stomach neoplasms Lactotransferrin 5-aza-CdR DNA methylation CpG island |
