摘要
目的检测并验证miR-134及miR-17～92基因簇在紫杉醇耐药与敏感卵巢癌细胞中的差异表达并探讨两者参与卵巢癌耐药的可能机制。方法2010年10月至2012年12月于中国医科大学附属盛京医院，用Affymetrix GeneChip miRNA芯片对紫杉醇耐药与敏感卵巢癌细胞miRNA基因表达谱进行分析。选取差异表达明显的miR-134和miR-17～92基因簇，采用Real-timePCR方法进一步验证芯片结果。应用生物信息学分析软件预测miR-134和miR-17～92基因簇各自潜在的靶基因并通过WesternBlot对部分靶基因进行验证。结果与敏感卵巢癌SKOV3细胞相比，耐药卵巢癌SKOV3-TR30细胞中miR-134低表达，miR-17～92基因簇高表达。软件预测得到miR-134基因簇潜在靶基因c-Myc，MRP1／ABCCl等共128个，软件预测得到miR-17～92基因簇潜在靶基因BIM，PTEN，ABCAl等共30个。与敏感卵巢癌SKOV3细胞相比，耐药卵巢癌SKOV3-TR30细胞中miR-134潜在靶基因c-Myc蛋白高表达；miR-17～92基因簇潜在靶基因BIM蛋白低表达（P均〈0．05），PTEN蛋白虽表达增高但差异无统计学意义。结论MicroRNA表达谱基因芯片可筛查出紫杉醇耐药与敏感卵巢癌细胞的差异表达基因。低表达miR-134和高表达的miR-17～92基因簇可能分别通过调节其潜在靶基因c-Myc及BIM蛋白表达参与卵巢癌化疗耐药。
Objective To detect the differentially miRNA expression in paclitaxel sensitive and chemoresistant ovarian cancer cell lines, and to explore the molecular mechanism of drug resistance of ovarian cancer. Methods The expression profile of miRNA were detected with miRNA microarray chip hybridization and the compared array results were further validated by Real-time PCR and Western Blot. Potential miRNA targets were predicted by bioinformatic software. Results Compared with SKOV3 cell line, miR17 -92 gene cluster with high expression while miRNA134 cluster with low expression was found in SKOV3-TR30 cells. It can be predicted by bioinformatic software that miR17 -92 gene cluster has 30 target genes such as BIM,PTEN,ABCA1 ,C10orf46,CD4, while miRNA134 cluster has 128 target genes such as MRP1/ABCC1. Western Blot showed that compared with SKOV3 ceils, BIM expression was decreased in SKOV3- TR30 cells,while c-Myc was in high expression in SKOV3-TR30 cells. Conclusion The differentially miRNA expression profiles in SKOV3 and SKOV3-TR30 can be obtained by miRNA microarray chip technology. MiR-17 -92 cluster and miR-134 cluster may be involved in drug resistance of ovarian cancer by regulating their target genes,which provide a theoretical basis for the study of ovarian cancer chemoresistance.
出处
《中国实用妇科与产科杂志》
CAS
CSCD
北大核心
2014年第2期118-122,共5页
Chinese Journal of Practical Gynecology and Obstetrics
基金
国家自然科学基金(30973189)
辽宁省自然科学基金(2013021040)
