中华泌尿外科杂志
中華泌尿外科雜誌
중화비뇨외과잡지
CHINESE JOURNAL OF UROLOGY
2014年
3期
222-225
,共4页
张永海%陈凌武%徐庆春%林伟强%黄月英%黄洪%张源锋%刘国元
張永海%陳凌武%徐慶春%林偉彊%黃月英%黃洪%張源鋒%劉國元
장영해%진릉무%서경춘%림위강%황월영%황홍%장원봉%류국원
膀胱癌%微小RNA%芯片%实时定量PCR%血浆
膀胱癌%微小RNA%芯片%實時定量PCR%血漿
방광암%미소RNA%심편%실시정량PCR%혈장
Bladder cancer%microRNA%Microarray%Real-time PCR%Plasma
目的 寻找并鉴定在膀胱癌和正常膀胱组织,及患者血浆中差异表达的微小RNA(miRNA).方法 2010年10月至201 1年1月收集24例膀胱尿路上皮癌(T1～4期)患者的新鲜肿瘤组织及血浆,其中非肌层浸润性和肌层浸润性膀胱尿路上皮癌组织各12例;12例良性前列腺增生患者的正常膀胱组织及血浆作为对照组.用Trizol试剂提取总RNA,用Nanodrop 2000及变性琼脂糖凝胶电泳进行RNA检测,用标记酶Hy3TM荧光基团标记RNA的探针和miRCURYTM芯片杂交,用GenePix 4000B芯片扫描仪及GenePix pro V6.0进行图像采集和数据分析,对差异表达的miRNA进行检测,用实时定量逆转录聚合酶链反应(real-time quantitative PCR,qPCR)对部分差异表达的miRNA进行验证.结果 与正常膀胱组织比较,在非肌层浸润性膀胱癌组织中,有77个miRNA表达上调,72个miRNA表达下调;在肌层浸润性膀胱癌组织中,42个miRNA表达上调,104个miRNA表达下调.qPCR测定miR-720和miR-191的表达水平与基因芯片结果一致.与对照组比较,膀胱癌血浆样本中有19个miRNA表达上调,9个miRNA表达下调.qPCR测定miR-10b、miR-622、miR-634和miR-483-3p的表达水平变化趋势与基因芯片结果一致.结论 在非肌层浸润性和肌层浸润性膀胱尿路上皮癌组织及血浆中miRNA的差异表达普遍存在,可作为膀胱癌的潜在临床诊断标志物.
目的 尋找併鑒定在膀胱癌和正常膀胱組織,及患者血漿中差異錶達的微小RNA(miRNA).方法 2010年10月至201 1年1月收集24例膀胱尿路上皮癌(T1～4期)患者的新鮮腫瘤組織及血漿,其中非肌層浸潤性和肌層浸潤性膀胱尿路上皮癌組織各12例;12例良性前列腺增生患者的正常膀胱組織及血漿作為對照組.用Trizol試劑提取總RNA,用Nanodrop 2000及變性瓊脂糖凝膠電泳進行RNA檢測,用標記酶Hy3TM熒光基糰標記RNA的探針和miRCURYTM芯片雜交,用GenePix 4000B芯片掃描儀及GenePix pro V6.0進行圖像採集和數據分析,對差異錶達的miRNA進行檢測,用實時定量逆轉錄聚閤酶鏈反應(real-time quantitative PCR,qPCR)對部分差異錶達的miRNA進行驗證.結果 與正常膀胱組織比較,在非肌層浸潤性膀胱癌組織中,有77箇miRNA錶達上調,72箇miRNA錶達下調;在肌層浸潤性膀胱癌組織中,42箇miRNA錶達上調,104箇miRNA錶達下調.qPCR測定miR-720和miR-191的錶達水平與基因芯片結果一緻.與對照組比較,膀胱癌血漿樣本中有19箇miRNA錶達上調,9箇miRNA錶達下調.qPCR測定miR-10b、miR-622、miR-634和miR-483-3p的錶達水平變化趨勢與基因芯片結果一緻.結論 在非肌層浸潤性和肌層浸潤性膀胱尿路上皮癌組織及血漿中miRNA的差異錶達普遍存在,可作為膀胱癌的潛在臨床診斷標誌物.
목적 심조병감정재방광암화정상방광조직,급환자혈장중차이표체적미소RNA(miRNA).방법 2010년10월지201 1년1월수집24례방광뇨로상피암(T1～4기)환자적신선종류조직급혈장,기중비기층침윤성화기층침윤성방광뇨로상피암조직각12례;12례량성전렬선증생환자적정상방광조직급혈장작위대조조.용Trizol시제제취총RNA,용Nanodrop 2000급변성경지당응효전영진행RNA검측,용표기매Hy3TM형광기단표기RNA적탐침화miRCURYTM심편잡교,용GenePix 4000B심편소묘의급GenePix pro V6.0진행도상채집화수거분석,대차이표체적miRNA진행검측,용실시정량역전록취합매련반응(real-time quantitative PCR,qPCR)대부분차이표체적miRNA진행험증.결과 여정상방광조직비교,재비기층침윤성방광암조직중,유77개miRNA표체상조,72개miRNA표체하조;재기층침윤성방광암조직중,42개miRNA표체상조,104개miRNA표체하조.qPCR측정miR-720화miR-191적표체수평여기인심편결과일치.여대조조비교,방광암혈장양본중유19개miRNA표체상조,9개miRNA표체하조.qPCR측정miR-10b、miR-622、miR-634화miR-483-3p적표체수평변화추세여기인심편결과일치.결론 재비기층침윤성화기층침윤성방광뇨로상피암조직급혈장중miRNA적차이표체보편존재,가작위방광암적잠재림상진단표지물.
Objective To identify the differentially expressed microRNA (miRNA) in bladder cancer tissue and plasma.Methods 24 bladder urothelial carcinoma (BUC) fresh tissues and 12 normal bladder mucosa tissue were stored in liquid nitrogen.Total RNA was extracted by using Trizol reagents,purified and tested by denaturing agarose gel electrophoresis and NanoDrop 2000.The purified RNA were labeled by Hy3TM fluorescent label then hybridized with miRCURYTM Array.MicroRNA array scanning and analysis were performed with the Axon GenePix 4000B microarray scanner and GenePix proV6.0.The scanned result was validated by using qPCR.Results 77 microRNA expressions were increased and 72 microRNA expressions were decreased in non-invasive bladder cancer compared to the normal tissue.42 microRNA expressions were increased and 104 microRNA expressions were decreased in invasive bladder cancer compared to the normal tissue.The expression levels of miR-720 and miR-191 were verified by qPCR.The expression of microRNA was different between cancer tissue and plasma.19 microRNA expressions were increased and 9 microRNA expressions were decreased in the plasma of patients with bladder cancer compared to the normal plasma.The different expression levels of miR-10b,miR-622,miR-634 and miR-483-3p were verified by qPCR.Conclusions microRNA differentiations occur in a tumor phenotype-specific manner and might be potential clinical diagnostic markers.
