SOX7基因在胰腺癌细胞株的表达及其启动子区甲基化状态的研究
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国家自然科学基金面上项目(30500492);江苏省自然科学基金(BK2006241)


The expression and promoter methylation status of SOX7 gene in pancreatic cancer cell line
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    摘要:

    目的:检测SOX7基因在胰腺癌细胞株中的表达及其启动子区的甲基化状态,初步探讨两者之间的关系-方法:采用RT-PCR检测SOX7基因在胰腺癌细胞株BXPC-3-CFPAC-1-PANC-1和SW1990中的表达水平-重亚硫酸盐测序PCR(bisulfite sequencing PCR,BSP)和结合重亚硫酸盐的限制性内切酶法 (combined bisulfite restriction analysis,COBRA)检测启动子区域甲基化状态-采用去甲基化药物5-氮杂胞苷(5-aza-2-adeoxycytidine,5-aza-dC)处理各细胞株后,检测SOX7基因的mRNA表达和甲基化状态变化-结果:在胰腺癌细胞株BXPC-3-CFPAC-1中SOX7基因的mRNA呈阳性表达,而在PANC-1和SW1990中SOX7基因表达沉默;与BXPC-3-CFPAC-1相比,PANC-1和SW1990中SOX7基因启动子区甲基化率较高-经过去甲基化处理后,PANC-1与SW1990中SOX7启动子区的甲基化率下降,基因重新表达-结论:胰腺癌细胞株PANC-1和SW1990中的SOX7基因表达与启动子区甲基化状态有关,启动子区的高甲基化导致PANC-1- SW1990中SOX7基因的表达沉默-

    Abstract:

    Objective:To explore whether aberrant methylation is a inhibition factor to transcriptional inactivation of SOX7 in cancer cell lines by investigation of the expression and methylation status of SOX7 gene in human pancreatic cancer cell lines. Methods:RT-PCR method was used to explore mRNA expression of the SOX7gene. Bisulfite sequencing PCR(BSP) and combined bisulfite restriction analysis(COBRA) were used to test promoter methylation of SOX7 gene in BXPC-3,CFPAC-1,PANC-1 and SW1990. Re-testing of these two indicators after the treatment of 5-aza-2-deoxycytidine(5-aza-dC). Results:SOX7 gene was obviously expressed in BXPC-3 and CFPAC-1 but silenced in PANC-1 and SW1990. The rate of methylation of CpG island of SOX7 in PANC-1 and SW1990 was much higher than the other two genes. After the treatment of 5-aza-dC in hypermethylation cell line PANC-1 and SW1990,the methylation rate of SOX7 gene promoter was decreased and the mRNA of SOX7 was re-expressed. Conclusion:This promoter hypemlethylation is correlated with SOX7 gene expression in pancreatic cancer cell line PANC-1 and SW1990 and plays a key role in SOX7 silencing.

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彭〓泉,蔡辉华,高文涛,钱祝银,苗〓毅. SOX7基因在胰腺癌细胞株的表达及其启动子区甲基化状态的研究[J].南京医科大学学报(自然科学版),2010,(6):752-755865

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  • 收稿日期:2010-02-23
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