应用组织芯片技术筛选脑膜瘤差异表达蛋白

目的 应用组织芯片技术在蛋白质水平探讨脑膜瘤恶性变机制.方法 挑选79例脑膜瘤样本制备组织成芯片,与MYC、MDM2、ARNT2、AR、ER、PR、KJ-67、P53、survivin、CD34和VEGF共计11种抗体进行免疫组化实验.整个实验设置阴性对照组芯片且性激素受体应用乳腺癌芯片作为阳性对照.应用SAS9.0软件处理数据,筛选不同病理级别间的差异表达抗原.结果 成功制备了组织芯片.免疫组化染色信号清晰.没有非特异性染色.数据处理结果显示:P53、AR、CD34、Ki-67和MYC在脑膜瘤不同病理级别间阳性表达量差异存在统计学意义(P<0.05).结论 组织芯片结合免疫组化技术是在蛋白质水平探讨脑膜瘤恶性变机制的科学方法.P53、CD34、Ki-67、AR和MYC在脑膜瘤中的表达与其恶性变机制有关.是脑膜瘤病理级别诊断的重要标记物.

Tissue microarray-based high-throughput screening of differentially expressed proteins as biomarkers in meningioma

Objective To investigate the mechanism responsible for the malignant progressionof meningiomas at the protein level using tissue microarray technique. Methods Twenty-twointracranial meningioma tissue microarrays were constructed, each containing the tissues of 42 benign, 18atypical, and 19 anaplastic meningiomas. Immunohistochcmistry of the microarrays was performed induplicate with the antibodies of MYC, ARNT2, MDM2, AR, ER, PR, Ki-67, P53, survivin, CD34 andVEGF, respectively. Negative control microarrays were used throughout the experiment and breast cancertissue microarrays were used as the positive controls for ER and PR staining. SAS9.0 solfware was usedfor grading of the expression levels of the biomarkers according to the WHO grades of meningiomas.Results For each antibody, the duplicate tissue microarrays yielded uniform staining results invisualization of the protein distributions in the cytoplasm and nuclei, and the negative controls displayedno positive staining. The p53, AR, ER, PR and Ki-67 proteins were found only in the cell nuclei, MDM2in both the cytoplasm and nuclei, and ARNT2, CD34 and VEGF in the cytoplasm only. The c-MYC andsurvivin proteins were found mainly in the cytoplasm, and in some instances in both the cytoplasm andcell nuclei. Immunohistochemical staining for p53, AR, CD34, Ki-67 and MYC proteins showed strongcorrelations to the degree of malignancy of the meningioma (P<0.05). Conclusions Tissue microarrayand immunohistochemical techniques provide an efficient means for screening the specific biomatkers ofmeningiomas. The expressions of p53, AR, CD34, Ki-67 and MYC proteins are involved in the malignantprogression of meningioma, and these proteins may serve as important biomarkers for meningiomagrading at the protein level.