应用量子点技术检测Bcl-2和P53在皮肤血管瘤中的表达

The Expression of Bcl-2 and P53 in Human Dermal Hemangiomas Detected by the Quantum Dot Technology

目的通过量子点技术检测Bcl-2和P53在增生期、退化期皮肤血管瘤及正常组织中的表达,进一步阐明Bcl-2和P53在皮肤血管瘤细胞增殖与凋亡中的作用。方法选取武汉大学人民医院病理科2007年至2010年皮肤毛细血管瘤存档蜡块38例(男性16例,女性22例),血管瘤周围正常皮肤组织5例。对所有标本进行常规苏木精-伊红染色,并使用免疫组织化学SP法检测增殖细胞核抗原(PCNA),根据Mulliken提出的分类标准并结合PCNA的表达情况将所有血管瘤组织分为增生期组和退化期组。然后采用免疫组织化学SP法以及量子点双染技术检测38例皮肤血管瘤及5例正常瘤旁皮肤组织中Bcl-2和P53蛋白及PCNA的表达水平;采用多光谱成像系统对量子点染色结果进行图像分析,并用SPSS 13.0软件对各组量子点染色分析后的数据行单因素方差分析和SNK(q)检验,检验水准α为0.05。结果①38例标本中,增生期血管瘤有24例,退化期血管瘤有14例。②免疫组织化学SP法显示,增生期血管瘤有Bcl-2和P53表达,瘤旁正常皮肤组织中和退化期血管瘤组织中Bcl-2和P53表达极弱或无表达。③量子点染色及统计分析显示,Bcl-2和P53在增生期血管瘤中的表达显著高于退化期血管瘤和瘤旁正常皮肤组织(均P<0.05),而退化期血管瘤和瘤旁正常皮肤组织之间Bcl-2和P53的表达差异无统计学意义。结论 Bcl-2和P53在皮肤血管瘤的发生发展中发挥重要作用。Bcl-2可能是通过抑制血管瘤增生期内皮细胞的凋亡,使其增殖和凋亡失衡;P53可能促进内皮细胞的增殖,使血管内皮细胞大量生成。

Objective To detect the expression of Bcl-2 and P53 in different phases of hemangioma by using high-sensitive quantum dot dyeing method in order to better understand the role of Bcl-2 and P53 in the proliferation and apoptosis of heman- giomas cells. Methods This study involved 38 paraffin-embedded hemangioma specimens（involving 16 males and 22 females） and 5 paraffin-embedded normal skin tissues which were archived in the Department of Pathology, Renmin Hospital of Wuhan University between 2007 to 2010. Immunohistochemistry,regular HE staining and quantum dot dyeing were used to detect the expression of proliferating cell nuclear antigen（PCNA）, Bcl-2 and P53 in hemangioma and normal skin tissues. All the hemangi- oma specimens were classified into the proliferation or the involution stage according to the Mulliken＇ s standard and the expres- sion of PCNA. The results of the quantum dot dyeing were analyzed by multi-spectral imaging systems. Analysis of variance （ANOVA） and SNK（q） test were used with SPSS 13.0 to analyze the data from the quantum dot dyeing（a=0. 05）. Results There were 24 cases of hemangioma at proliferation stage and 14 at involution stage in 38 paraffin-embedded hemangioma speci- mens. Immunohistochemical results indicated that the expression levels of Bcl-2 and P53 were significantly higher in hemangio- ma tissues at proliferation stage than at involution stage. The results from quantum dot dyeing showed that the expression of Bcl-2 and P53 in the proliferating hemangiomas was significantly higher than that in involuting ones（P〈0.05） and in normal tissues（P〈0.05）. No significant difference in the Bcl-2 and P53 expression was found between the involuting hemangiomas and normal tissues. Conclusion Bcl-2 may cause the imbalance of proliferation and apoptosis in endothelial cells by inhibiting the endothelial cell apoptosis and P53 may promote the proliferation of endothelial cells in proliferating dermal hemangioma and contribute to the generation of a large number of vascular endothelial cells.