miR-34启动子甲基化在胃癌发病过程中的作用研究
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摘要
目的探讨胃癌组织中miR-34基因启动子甲基化状态对miR-34的表达影响及其与患者预后的相关性。方法采用实时荧光定量PCR法检测50例胃癌组织及癌旁组织中成熟miR-34的表达。采用甲基化特异性PCR法检测miR-34基因启动子的甲基化状态,运用统计学分析miR-34基因启动子甲基化状态与miR-34表达量之间的关系,并随访患者复发和转移情况。结果胃癌组织中成熟miR-34的表达水平比癌旁组织明显降低(P<0.05)。miR-34甲基化水平与淋巴结转移有相关性(P<0.05)。miR-34基因启动子非甲基化组的成熟miR-34表达水平明显高于甲基化阳性单一模式组和甲基化阳性混合模式组(P<0.05)。与非甲基化组和甲基化阳性混合模式组相比,miR-34基因甲基化阳性组的胃癌患者复发和转移率明显升高(P<0.05)。结论在胃癌患者中,miR-34基因启动子甲基化可导致miR-34表达下调。miR-34基因启动子的甲基化状态与胃癌的复发与转移明显相关。
Objective To investigate the effect of miR-34 gene promoter methylation status on the expression of miR-34 in gastric cancer and its correlation with prognosis. Methods The expression of mature miR-34 in 50 gastric cancer tissues and adjacent tissues was detected by real-time fluorescent quantitative PCR. The methylation status of the miR-34 gene promoter was detected by methylation-specific PCR. The relationship between miR-34 gene promoter methylation status and miR-34 expression was analyzed and the patients were followed up for recurrence and metastasis. Results The expression of mature miR-34 in gastric cancer tissues was significantly lower than that in the control group(P<0.05). miR-34 expression was correlated with lymph node metastasis(P<0.05). The expression level of mature miR-34 in non-methylated group was significantly higher than that in methylated positive single mode group and methylated positive mixed mode group(all P<0.05). Compared with non-methylated group and methylated positive mixed group,the recurrence and metastasis rate of gastric cancer patients with miR-34 methylation positive were significantly higher(P<0.05). Conclusion Methylation of miR-34 gene may lead to the down-regulation of miR-34 expression in some gastric cancer patients. The methylation status of miR-34 is associated with the recurrence and metastasis rate of gastric cancer.
Objective To investigate the effect of miR-34 gene promoter methylation status on the expression of miR-34 in gastric cancer and its correlation with prognosis. Methods The expression of mature miR-34 in 50 gastric cancer tissues and adjacent tissues was detected by real-time fluorescent quantitative PCR. The methylation status of the miR-34 gene promoter was detected by methylation-specific PCR. The relationship between miR-34 gene promoter methylation status and miR-34 expression was analyzed and the patients were followed up for recurrence and metastasis. Results The expression of mature miR-34 in gastric cancer tissues was significantly lower than that in the control group(P<0.05). miR-34 expression was correlated with lymph node metastasis(P<0.05). The expression level of mature miR-34 in non-methylated group was significantly higher than that in methylated positive single mode group and methylated positive mixed mode group(all P<0.05). Compared with non-methylated group and methylated positive mixed group,the recurrence and metastasis rate of gastric cancer patients with miR-34 methylation positive were significantly higher(P<0.05). Conclusion Methylation of miR-34 gene may lead to the down-regulation of miR-34 expression in some gastric cancer patients. The methylation status of miR-34 is associated with the recurrence and metastasis rate of gastric cancer.
引文
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[14]殷帅,徐仁芳,何小舟,等.肾透明细胞癌组织中miR-34a和miR-34b/c基因CpG岛甲基化状态的变化及意义[J].实用临床医学,2013,14(6):5-8.
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[16]Siemens H,Neumann J,Jackstadt R,et al.Detection of miR-34a promoter methylation in combination with elevated expression of c-Met and beta-catenin predicts distant metastasis of colon cancer[J].Clin Cancer Res,2013,19(3):710-720.
[17]Schmid G,Notaro S,Reimer D,et al.Expression and promotor hypermethylation of miR-34a in the various histological subtypes of ovarian cancer[J].BMC Cancer,2016,16:102-130.
[2]Moutinho C,Esteller M.MicroRNAs and epigenetics[J].Adv Cancer Res,2017,135:189-220.
[3]Kim YH,Lee WK,Lee EB,et al.Combined effect of metastasis-related microRNA,miR-34 and miR-124family,methylation on prognosis of non-small-cell lung cancer[J].Clin Lung Cancer,2017,18(1):e13-e20.
[4]Vogt M,Munding J,Grüner M,et al.Frequent concomitant inactivation of miR-34a and miR-34b/c by CpG methylation in colorectal,pancreatic,mammary,ovarian,urothelial,and renal cell carcinomas and soft tissue sarcomas[J].Virchows Arch,2011,458(3):313-322.
[5]邓晓晶,邓敏,宋育林,等.miRNA-34a对人胃癌SGC-7901细胞增殖、凋亡的影响[J].安徽医科大学学报,2015,50(8):1090-1094.
[6]Mahul BA,Stephen E,Frederick LG,et al.AJCCcancer staging manual[M].8th ed.New York:Springer,2016.
[7]Timp W,Feinberg AP.Cancer as adysregulated epigenome allowing cellular growth advantage at the expense of the host[J].Nat Rev Cancer,2013,13(7):497-510.
[8]Ramassone A,Pagotto S,Veronese A,et al.Epigenetics and microRNAs in cancer[J].Int J Mol Sci,2018,19(2):E459.
[9]Slaby O,Laga R,Sedlacek O,et al.Therapeutic targeting of non-coding RNAs in cancer[J].Biochem J,2017,474(24):4219-4251.
[10]Kim NH,Cha YH,Kang SE,et al.p53 regulates nuclear GSK-3 levels through miR-34-mediated Axin2suppression in colorectal cancer cells[J].CellCycle,2013,12(10):1578-1587.
[11]Cha YH,Kim NH,Park C,et al.MiRNA-34 intrinsically links p53 tumor suppressor and Wnt signaling[J].Cell Cycle,2012,11(7):1273-1281.
[12]Vogt M,Munding J,Grüner M,et al.Frequent concomitant inactivation of miR-34a and miR-34b/c by CpG methylation in colorectal,pancreatic,mammary,ovarian,urothelial,and renal cell carcinomas and soft tissue sarcomas[J].Virchows Arch,2011,458(3):313-322.
[13]邵佳,何爱琴,张玉泉,等.miR-34甲基化与卵巢癌临床病理特征及预后的关系[J].江苏大学学报(医学版),2014,24(6):513-516,419.
[14]殷帅,徐仁芳,何小舟,等.肾透明细胞癌组织中miR-34a和miR-34b/c基因CpG岛甲基化状态的变化及意义[J].实用临床医学,2013,14(6):5-8.
[15]Suzuki H,Yamamoto E,Nojima M,et al.Methylation-associated silencing of microRNA-34b/c in gastric cancer and its involvement in an epigenetic field defect[J].Carcinogenesis,2010,31(12):2066-2073.
[16]Siemens H,Neumann J,Jackstadt R,et al.Detection of miR-34a promoter methylation in combination with elevated expression of c-Met and beta-catenin predicts distant metastasis of colon cancer[J].Clin Cancer Res,2013,19(3):710-720.
[17]Schmid G,Notaro S,Reimer D,et al.Expression and promotor hypermethylation of miR-34a in the various histological subtypes of ovarian cancer[J].BMC Cancer,2016,16:102-130.