促胃液素高表达大肠癌组织中microRNA表达差异分析
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皖南医学院第一附属医院,皖南医学院第一附属医院,皖南医学院第一附属医院

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安徽省自然科学基金(1408085MH148),皖南医学院弋矶山医院人才引进基金(YR201406),皖南医学院自然科学基金(WK2012zf02)


Profiling analysis of differential expression of microRNA in colorectal cancer tissues with high expression of gastrin
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Supported by Natural Science Foundation of Anhui Province (1408085MH148), Talent Introduction Science Fund of Yijishan Hospital (YR201406), and Natural Science Foundation of Wannan Medical College (WK2012zf02)

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    摘要:

    目的 运用基因芯片技术筛选促胃液素(GAS)高表达的大肠癌组织中差异表达的微RNA (miRNA)。方法 运用酶联免疫吸附实验(ELISA)检测71例大肠癌患者癌组织标本中GAS的含量,分析大肠癌组织中GAS表达水平与临床病理特征的关系。选择GAS表达强阳性(高表达组)与阴性(对照组)的患者各4例,对其癌组织进行miRNA芯片分析,并采用qPCR验证差异表达的miRNA。结果 GAS含量≥50.00 pg/g(阳性表达)的患者17例(23.9%),其余54例(76.1%) GAS表达阴性。癌组织中GAS表达水平与其分化程度、Dukes分期及组织学类型有关(P均<0.05 )。GAS高表达组(GAS含量>200.00 pg/g)中与对照组癌组织相比发生显著变化的miRNA共236个,与对照组比较,在高表达组中表达上调的miRNA共159个,表达下调的miRNA的数目为77个。筛选出GAS高表达组中的3倍以上表达差异的miRNA 24个,其中上调17个、下调7个;分别选择表达上调和下调的miRNA各3个进行qPCR验证,验证结果与芯片分析结果基本一致。结论 GAS高表达大肠癌组织中miRNA的表达存在显著变化,差异表达miRNA可能是治疗GAS高表达大肠癌的潜在靶点。

    Abstract:

    Objective To screen the differentially expressed microRNAs (miRNAs) in colorectal cancer tissues with high expression of gastrin (GAS) by gene chip technique. Methods The level of GAS in tumor tissues from 71 patients with colorectal cancer was detected by enzyme linked immumosorbent assay (ELISA), and the relationship between positive expression of GAS and clinicopathological features was analyzed. Human colorectal cancer tissues with high positive GAS expression (high-expression group, n=4) or negative GAS expression (control group, n=4) were analyzed by miRNA microarray, and the selected differentially expressed miRNAs were validated by qPCR. Results There were 17 cases (23.9%) with GAS content ≥50.00 pg/g (positive GAS expression), while the remaining 54 cases (76.1%) had negative GAS expression. The expression level of GAS was related to the degree of differentiation, Dukes stage and histological type (P<0.05). There were 236 miRNAs had significant differential expression in the high-expression group (GAS content >200.00 pg/g) compared with the control group, of which 159 miRNAs were up-regulated and 77 were down-regulated. A total of 24 miRNAs were screened out, which differentially expressed more than 3 folds in the high-expression group, with 17 up-regulated and 7 down-regulated. Three up-regulated or down-regulated miRNAs were selected for qPCR verification, and the results were consistent with the microarray analysis. Conclusion The miRNAs are differentially expressed in the colorectal cancer tissues with high expression of GAS, and they would be potential targets for the treatment of the colorectal cancer with high GAS expression.

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  • 收稿日期:2017-05-10
  • 最后修改日期:2017-06-10
  • 录用日期:2017-06-20
  • 在线发布日期: 2017-06-28
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