胃癌冰冻组织特定区域的微阵列制作及其特点分析

Construction and Characterization of Frozen Tissue Microarray with Defined Histological Regions of Gastric Specimens

背景与目的:目前癌症基因组学研究(CancerGenomicProjects,CGP)急需建立一种与大量候选基因再筛查和功能分析相配套的高通量下游检测技术。本研究旨在建立冰冻组织特定区域的定点捕获和组织阵列制作技术,以便在相同实验条件下高效检测各类胃粘膜病变的基因表达特点。方法:使用自制的组织微阵列制作装置,在0.7cm2的最佳切割温度复合物(optimalcuttingtemperaturecompound,OCT)载体上制作含30个组织点的胃癌/粘膜冰冻组织特定区域的微阵列。采用组织病理学和针对Caspase-3的免疫组织化学染色(immunohistochemistry,IHC)以及mRNA原位杂交技术(mRNA-insituhybridization,RISH),对组织微阵列的特点加以鉴定。结果:组织学观察显示,定点捕获的精确度较高;各组织点较好地保持了原有的形态。IHC和RISH的结果表明,Caspase-3在非癌胃粘膜组织、癌前病变组织和胃癌组织中的检出率分别为100.0%,66.7%,20.0%和90.0%,60.0%,15.4%。除因脱片和卷边所致的组织点不匹配外,两者结果的吻合度为100%。结论:本研究建立的冰冻组织微阵列技术使高效而准确地分析大量冰冻标本的组织学和细胞/分子遗传学特点成为可能,它在包括癌症基因组学在内的肿瘤相关基因筛选和功能分析中有着广泛的应用前景。

BACKGROUND &OBJECTIVE:In the g enome-wide investig ation of cancers ,hig h-throug hput downst ream approach is in urg ent need to val idate the candidates of cancer associated g enes screened by cDNA microarrays.To meet this need,frozen tissue microarrays were constructed using defined histolog ical reg ions of g astric tis sues and their g eneral features were characterized by multiple paramete rs.METHODS :By the use of self-desig ned tissue arraying system,0.7mm tissu e spots were cored from defined histo log ical reg ions of g astric tissues and order ly filled into the array-recipient O CT(optimal cutting temperature compound)block in the density of 30spots /0.7c m 2 or 49spots /cm 2 .The spots of tissue array was charac terized histolog ically,and then by the methods of immunohistochemis try(IHC)and mRNA-in situ hybridization(RISH)for casepase-3.RESULTS :The spot sampling was performed accurately,and the morpholog y of mo st tissue spots was kept well after mu ltiple steps of processing .Immunohistoch emical staining revealed that casep ase-3was produced in 100.0%of noncancero us g astric mucosa,66.7%of neoplast ic mucosa and 20.0%in cancer tissues.T he results of RISH were well coincide d with that of ICC in the rates of 90.0%,60.0%,and 15.4%.CONCLUSION:The frozen tissue array described in this study allows rapid and precise molecular analyses of various g astric lesions in the same experimental conditions .It would be a novel downstream tool in g enome-wide study of g astric cancers both in screening and functional elucidation of candidate g astric cancer associated g enes.