Sun Yuan, Kong Qing-You, Chen Xiao-Yan, Wang Xiao-Wei, Liu Jia, Li Hong
Cancer Institute, Dalian Medical University, Dalian, Liaoning, 116027, PR China.
Ai Zheng. 2003 Sep;22(9):1005-8.
BACKGROUND & OBJECTIVE: In the genome-wide investigation of cancers, high-throughput downstream approach is in urgent need to validate the candidates of cancer associated genes screened by cDNA microarrays. To meet this need, frozen tissue microarrays were constructed using defined histological regions of gastric tissues and their general features were characterized by multiple parameters.
By the use of self-designed tissue arraying system, 0.7 mm tissue spots were cored from defined histological regions of gastric tissues and orderly filled into the array-recipient OCT(optimal cutting temperature compound) block in the density of 30 spots/0.7 cm(2) or 49 spots/cm(2). The spots of tissue array was characterized histologically, and then by the methods of immunohistochemistry(IHC) and mRNA-in situ hybridization(RISH) for casepase-3.
The spot sampling was performed accurately, and the morphology of most tissue spots was kept well after multiple steps of processing. Immunohistochemical staining revealed that casepase-3 was produced in 100.0% of noncancerous gastric mucosa, 66.7% of neoplastic mucosa and 20.0% in cancer tissues. The results of RISH were well coincided with that of ICC in the rates of 90.0%, 60.0%, and 15.4%.
The frozen tissue array described in this study allows rapid and precise molecular analyses of various gastric lesions in the same experimental conditions. It would be a novel downstream tool in genome-wide study of gastric cancers both in screening and functional elucidation of candidate gastric cancer associated genes.
在癌症全基因组研究中，迫切需要高通量下游方法来验证通过cDNA微阵列筛选出的癌症相关基因候选物。为满足这一需求，利用胃组织特定的组织学区域构建了冷冻组织微阵列，并通过多个参数对其一般特征进行了表征。
使用自行设计的组织阵列系统，从胃组织特定的组织学区域钻取0.7mm的组织点，以30个点/0.7cm²或49个点/cm²的密度有序填充到阵列接受体OCT（最佳切割温度化合物）块中。对组织阵列的点进行组织学表征，然后通过免疫组织化学（IHC）和mRNA原位杂交（RISH）方法检测caspase-3。
点采样准确，经过多步处理后，大多数组织点的形态保持良好。免疫组织化学染色显示，caspase-3在100.0%的非癌胃黏膜、66.7%的肿瘤黏膜和20.0%的癌组织中产生。RISH结果与ICC结果的吻合率分别为90.0%、60.0%和15.4%。
本研究中描述的冷冻组织阵列能够在相同实验条件下对各种胃病变进行快速、精确的分子分析。它将成为胃癌全基因组研究中筛选和阐明候选胃癌相关基因功能的新型下游工具。
