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目的通过生物信息学与免疫组化实验相结合的方法,探讨新的卵巢癌发病相关microRNA(miRNA)。方法选取2004年1月至2006年3月间哈尔滨医科大学附属第一医院及哈尔滨医科大学附属第三医院收治的70例上皮性卵巢癌手术病例石蜡包埋样本,采用SP免疫组化法进行检测,设阴性对照组及阳性对照组,阳性反应观察采用双盲法。以Gene Ontology为功能组学背景,度量不同基因之间的功能相似性得分,根据得分综合排序,寻找卵巢癌风险miRNA。应用该方法对2 588个miRNA进行优化排序,通过免疫组化实验方法对风险miRNA在卵巢癌中调控位点进行证实。结果优化后,卵巢癌相关miRNA如miR-200a/b/c排在前列,排序前10位miRNA中的大部分都在正常组织和卵巢癌组织中呈差异表达。miR-125a调控的人表皮生长因子-2(HER-2)呈高表达趋势。HER-2表达水平与临床分期,病理分级及存活时间显著相关,差异有统计学意义(P<0.05)。结论 miR-125a通过调控HER-2影响卵巢癌的临床病理特征,可成为新的卵巢癌相关风险因子。
Objective To explore new ovarian cancer related microRNAs (miRNAs) through the combination of bioinformatics and immunohistochemistry. Methods Seventy patients with epithelial ovarian cancer who were admitted to the First Affiliated Hospital of Harbin Medical University and the Third Affiliated Hospital of Harbin Medical University between January 2004 and March 2006 were enrolled in this study. SP immunohistochemistry , Set negative control group and positive control group, the positive reaction was observed by double-blind method. Using Gene Ontology as a functional genomics background, the functional similarity score between different genes was measured. Based on the scores, the miRNAs were screened for the risk of ovarian cancer. This method was used to optimize the sequencing of 2 588 miRNAs, and the regulatory sites of risk miRNAs in ovarian cancer were confirmed by immunohistochemistry. Results After optimization, ovarian cancer-associated miRNAs such as miR-200a / b / c were in the forefront, and most of the top 10 miRNAs were differentially expressed in normal and ovarian cancers. miR-125a-regulated human epidermal growth factor-2 (HER-2) showed a high expression trend. The expression level of HER-2 was significantly correlated with clinical stage, pathological grade and survival time, the difference was statistically significant (P <0.05). Conclusion miR-125a may be a new risk factor of ovarian cancer by regulating the clinicopathological features of HER-2 in ovarian cancer.