摘要
目的探讨与胃肠道间质瘤(gastrointestinal stromal tumor,GIST)耐药相关的微小RNA(miRNA)。方法收集4例伊玛替尼耐药GIST和3例恶性GIST,运用Agilent人miRNA寡核苷酸基因芯片V3.0(955个已知miRNA)进行检测,选取耐药组与恶性组间3个差异表达最明显的miRNA,再应用RT-PCR技术在21个扩大样本中验证基因芯片筛选的结果。结果耐药组与恶性组相差倍数2倍以上(P<0.05),差异表达的miRNA有13个,上调者5个,下调者8个。其中差异最明显的3个分别为miR-15a、miR-1280、miR-320a,前者在耐药组中上调,后两者下调。RT-PCR技术在扩大样本中验证后发现miR-320a在两组中的差异与基因芯片的结果基本相符。结论通过miRNA芯片初步筛选和在扩大样本中的验证,发现miR-320a在耐药组与恶性组之间表达差异有显著性,提示miR-320a可能与GIST在甲磺酸伊玛替尼治疗过程中的耐药性有关。
Purpose Imatinib-resistance occurred in some patients with gastrointestinal stromal tumor （GIST） during the course of treatment. In this study, we preliminarily investigate the related miRNA in imatinib resistant GIST. Methods Seven GIST samples including imatinib-resistant group （ n = 4 ） and malignant group （ n = 3 ） were collected based on the clinincal and pathological charac- teristics. Total RNA was extracted, followed by miRNA microarray analysis to screen the differentially expressed miRNAs. Three miR- NAs presented with the most significantly expression were chosen for further validation by real-time PCR in 21 additional GISTs. Re- suits Thirteen miRNAs were revealed significantly differentially expression with P 〈 0. 05 and a fold change 〉 2 by comparison of imatinib-resistant group versus malignant group. Of them, five miRNAs were up-regulated and eight were down-regulated in imatinib- resistant group. Three miRNAs （miR-15a, miR-1280 and miR-320a） were the most significantly and differentially expressed between two groups. The differential expression of miR-320a was subsequently confirmed with concordance by real-time PCR. ConcLusions The differential miRNA expression profiles between two groups are revealed by miRNA microarray assay, and confirmed by real-time PCR. Among differentially expressed miRNAs, miR-320a might be related to imatinib-resistant GIST.
出处
《临床与实验病理学杂志》
CAS
CSCD
北大核心
2014年第2期123-127,共5页
Chinese Journal of Clinical and Experimental Pathology
基金
国家自然科学基金青年基金(81101809)
上海市卫生局科研课题(2009020)
