Promoter methylation regulates tazarotene-induced gene-1 expression in human pancreatic cancer
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摘要: 目的 研究他扎罗汀诱导基因1(tazarotene-induced gene-1,TIG1)在胰腺癌中的表达情况及启动子在胰腺癌细胞中的甲基化状态。 方法 收集67组胰腺癌和对应癌旁组织,用SP法进行TIG1蛋白检查并分析TIG1在癌、癌旁组织的表达差异。培养胰腺癌细胞株PANC1、BxPC3和SW1990,提取细胞总DNA,甲基化修饰后亚硫酸盐测序PCR(BSP)分析启动子CpG岛甲基化状态;筛选细胞总RNA,用实时定量PCR(real-time PCR)依次测各细胞株、经5-杂氮-2-脱氧胞嘧啶(5-aza-dC)干预前后TIG1 mRNA的表达差异。 结果 TIG1在癌组织中的表达低于癌旁组织(P<0.05)。TIG1的表达与胰腺癌的大小、分期无相关性,但与部位存在相关性(P<0.05)。TIG1在BxPC3和SW1990细胞中存在高甲基化状态,分别为(92±2)%和(86±2)%,在PANC1细胞甲基化率为(40±2)%。TIG1甲基化的BxPC3和SW1990细胞经5-aza-dC处理后TIG1表达明显上调(P<0.05),而甲基化的PANC1细胞经同样处理后TIG1的表达变化不明显。 结论 癌组织中TIG1的表达显著低于癌旁组织。不同部位胰腺癌TIG1的表达存在差异。TIG1基因在BxPC3和SW1990胰腺癌细胞中的低表达与甲基化有关。TIG1基因甲基化可能是调控胰腺癌细胞TIG1表达的重要分子机制之一。
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关键词:
- 胰腺癌 /
- 他扎罗汀诱导基因1 /
- 高甲基化 /
- 5-杂氮-2-脱氧胞嘧啶
Abstract: Objective To investigate the expression of tazarotene-induced gene-1 (TIG1) in pancreatic cancer and the methylation status of TIG1 promoter in pancreatic cancer cells. Methods Total 67 cases of pancreatic carcinoma and 67 cases of paired adjacent tissue were collected as the object of study. TIG1 protein was detected by immunohistochemical SP method to analyze the difference of expression of TIG1 in cancer tissues and paired adjacent tissue. Methylation status of TIG1 promoter in pancreatic cancer cell lines PANC1, BxPC3 and SW1990 were examined by Bisulfite sequencing PCR (BSP). The levels of TIG1 mRNA in these cells were identified by RT-PCR. PANC1 BxPC3 and SW1990 cells were treated with the demethylation agent, 5-aza-dC, at different concentrations, and the changes of TIG1 mRNA in these cells were examined by RT-PCR. Results The expression of TIG1 in cancer tissues was lower than that in paired adjacent tissue (P<0.05), and the expression of TIG1 was not correlated with size stage and lymph node metastasis of pancreatic cancer, but there was correlation with location (P<0.05). (92±2)% and (86±2)% of TIG1 CpG sites were methylated in BxPC3 and SW1990 cells respectively. While the ratio was (40±2)% in PANC1 cells. Decreased TIG1 mRNA can be restored in BxPC3 and SW1990 after treatment with 5-aza-dC, while there was no significant change in PANC1 cells. Conclusion The expression of TIG1 in cancer tissues is lower than that in paired adjacent tissue. There is difference in the expression of TIG1 in different parts of pancreatic carcinoma. Decreased TIG1 in BxPC3 and SW1990 is significantly correlated with promoter hypermethylation, suggesting that promoter hypermethylation may play an important role in the regulation of TIG1.-
Key words:
- Pancreatic cancer /
- Tazarotene-induced gene-1 /
- Hypermethylation /
- 5-aza-dC
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