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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≥50.00pg/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>200.00pg/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。
Objective To screen differentially expressed microRNAs (miRNAs) in colorectal cancer tissues highly expressed by gastrin (GAS) using gene chip technology. Methods The expression of GAS in 71 specimens of colorectal cancer was detected by enzyme linked immunosorbent assay (ELISA), and the relationship between GAS expression and clinicopathological features was analyzed. Four patients with strong positive GAS expression (high expression group) and negative (control group) were selected and miRNA microarray analysis was performed on their cancer tissues. Differentially expressed miRNAs were verified by qPCR. Results Seventeen patients (23.9%) had GAS levels≥50.00pg / g (positive expression), while the remaining 54 patients (76.1%) had negative GAS expression. The expression of GAS in cancer tissues was correlated with the degree of differentiation, Dukes stage and histological type (all P <0.05). A total of 236 miRNAs were found in GAS overexpression group (GAS> 200.00pg / g), which showed significant changes compared with the control group. There were 159 miRNAs that were up-regulated in the high-expression group compared with the control group The number of miRNAs is 77. Twenty-four miRNAs with more than 3-fold higher expression in GAS-overexpressing group were screened, 17 of them were up-regulated and 7 were down-regulated. Three miRNAs that were upregulated and downregulated were selected and verified by qPCR, respectively. . Conclusion The expression of miRNA in GAS overexpressing colorectal cancer has significant changes. Differentially expressing miRNA may be a potential target for the treatment of GAS overexpressing colorectal cancer.