miR-10a-5p基因甲基化对卵巢癌铂类耐药的影响

发布时间：2018-03-04 01:03

本文选题：miR-a-p　切入点：甲基化　出处：《中华肿瘤防治杂志》2017年06期 　论文类型：期刊论文

【摘要】：目的卵巢癌是死亡率最高的妇科肿瘤,较强的化疗耐药性是其预后差的主要原因之一,为了阐明卵巢癌对铂类药物的耐药机制,本研究探讨miRNA基因的甲基化水平对卵巢癌铂类耐药的影响。方法将卵巢癌组织分为敏感组和耐药组,每组各3例;采用基因芯片技术,对比分析了两组微RNA(microRNA,miRNA)的表达差异;采用实时荧光定量PCR,分别在6例敏感和3例耐药组织、铂类药物敏感(CoC1)和耐药的卵巢癌细胞系(CoC1/DDP),检测了候选miRNA的表达差异;应用Massarray技术,检测敏感组织(15例)与耐药组织(6例)中miRNA基因启动子的甲基化差异;应用生物信息学分析,鉴定目标miRNA的潜在靶基因。结果以铂类药物敏感的卵巢癌组织样本为对照,利用基因芯片筛选,鉴定了6条在耐药组织样本中出现表达上调的miRNA(miR-493-3p、miR-10a-5p、miR-16-2-3p、miR-1248、miR-451a、miR-628-3p)和6条表达下调的miRNA(miR-509-3p、miR-1197、miR-376a-3p、miR-1273a、miR-550a-3p、miR-19b-3p)。组织验证发现,miR-509-3p、miR-493-3p、miR-10a-5p、miR-16-2-3p和miR-451a,与芯片结果一致;培养细胞研究发现,4条miRNA的表达调控方式与组织芯片结果一致,miR-10a-5p、miR-16-2-3p、miR-1248和miR-628-3p在耐药细胞系中高表达。进一步研究发现,与敏感肿瘤组织相比,耐药组织中miR-10a-5p基因启动子的甲基化水平出现显著降低,P=0.04。结合生物信息学预测HOXA1和USF2为miR-10a-5p与耐药相关的靶基因。结论与敏感组相比,耐药组miR-10a-5p基因启动子甲基化水平显著降低,miR-10a-5p表达升高,通过抑制HOXA1和USF2,抑制细胞凋亡,导致铂类化疗药物耐受。
[Abstract]:Ovarian cancer is the highest mortality rate of gynecologic tumors, strong resistance to chemotherapy is one of the main causes of poor prognosis in ovarian cancer, in order to clarify the mechanism of drug resistance to platinum drugs, this study investigated the effects of methylation level of miRNA gene of platinum resistance in ovarian carcinoma. Methods the ovarian cancer tissues were divided into sensitive group and resistant group, 3 cases in each group; using gene chip technology, comparative analysis of two groups of micro RNA (microRNA, miRNA) the difference of expression; using real-time fluorescence quantitative PCR, respectively in 6 cases and 3 cases of drug sensitive tissue, platinum sensitive ovarian cancer cell line (CoC1) and resistance (CoC1/DDP) detection. The differential expression of the candidate miRNA; the application of Massarray technology, detection of sensitive tissue (15 cases) and resistant tissues (6 cases) methylation difference in miRNA gene promoter; analysis by bioinformatics, potential target gene identification of target miRNA. The results of platinum Ovarian cancer tissue samples for drug sensitive control, screened by gene chip, 6 appeared in resistant tissue samples and up-regulated miRNA were identified (miR-493-3p, miR-10a-5p, miR-16-2-3p, miR-1248, miR-451a, miR-628-3p) and 6 downregulated miRNA (miR-509-3p, miR-1197, miR-376a-3p, miR-1273a, miR-550a-3p, miR-19b-3p). Results showed that miR-509-3p, miR-493-3p, miR-10a-5p, miR-16-2-3p and miR-451a, consistent with the microarray results; cell culture study found that the expression pattern of 4 miRNA were consistent with those of tissue microarray, miR-10a-5p, miR-16-2-3p, miR-1248 and miR-628-3p are highly expressed in resistant cells. Further studies showed that the tumor tissue compared with sensitive, methylation level miR-10a-5p resistant tissue gene promoter significantly decreased, P=0.04. combined forecasting of biological information of HOXA1 and USF2 for miR-10a-5p associated with drug resistance Conclusion: compared with the sensitive group, the methylation level of miR-10a-5p gene promoter is significantly decreased, the expression of miR-10a-5p is increased, and the inhibition of apoptosis by HOXA1 and USF2 can lead to platinum resistant chemotherapeutic drugs.

【作者单位】：北京大学第三医院妇产科;北京大学药学院天然药物及仿生药物国家重点实验室;
【基金】：2016年国家自然科学基金(面上项目81572765) 2013年国家自然科学基金(青年基金81202040)
【分类号】：R737.31

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