应用免疫组织化学方法评价组织芯片的可靠性

Evaluation of the reliability of tissue chip with immunohistochemistry by detecting ER and PR expression in human breast cancer

目的探讨组织芯片技术在免疫组化试验中的可靠性和提高组织芯片可信性的方法。方法选择82例人乳腺癌标本石蜡蜡块,利用传统制片技术和自行研制的专用器具分别制作常规石蜡切片和组织芯片,采用免疫组化技术检测雌激素受体(ER)、孕激素受体(PR)的表达。结果采用自行研制的专用器具制备组织芯片,所得样本的可分析率均在92.9%以上,且随着对同一标本取材数量的增加,其可分析标本率也明显上升。采用传统方法检测的82例乳腺癌标本中ER和PR的阳性率分别为61%和58.5%;采用组织芯片技术,一式一份取材,其阳性率分别为51.9%和50.0%,一式两份方式取材,其阳性率分别为53.8%和53.8%,采用一式三份方式取材,其阳性率分别为57.1%和60.7%,且三种不同取材方式的检测结果与传统制片法之间并无统计学差异(P>0.75)。结论在采用统一制备标准并保证组织芯片质量的前提下,挖取直径1.5mm的组织样本制备组织芯片,尽管组织芯片技术与传统方法检测的结果的一致率仍随着对同一标本取材次数的增加而提高,但是三种不同取材方式的检测结果之间并无统计学差异。因此,一式一份取材制备组织芯片完全可替代传统制片技术用于免疫组织化学乃至原位杂交、荧光原位杂交技术的研究,且可保证组织芯片的可信性和高通量特征。

Objective To explore the reliability of tissue chip （tissue microarray） and the methods to improve the reliability. Methods A total of 82 formalin-fixed, paraffin-embedded tissue blocks of breast cancer specimens were retrieved, along with their corresponding hematoxylin and eosin （H＆E）-stained slide, from the archives of the Department of Pathology, Second Hospital of Xi＇an Jiaotong University. Patent tissue micoarrayer was used to prepare tissue chip. Immunohistochemistry was performed to analyze the expression of estrogen receptor （ER） and progesterone receptor （PR） on the tissue chip. Results More than 92.9% tissue samples on the tissue chip were analyzed. The more core samples were punched from each original block, the more specimens could be analyzed. The positive rate of the expression of ER and PR on conventional tissue section were 61%and 58.5% respectively. When one core sample was punched from each original block, the positive rate of the expression of ER and PR on the tissue chip were 51.9%and 50.0% respectively. When two core samples were punched from each original block, the positive rate were 53.8% and 53.8% respectively. When three core samples were punched from each original block, the positive rate were 57.1% and 60.7% respectively. However, there was no statistic difference between the values using the above three methods（P〉0.75）. Conclusion With the same preparation standard and quality of tissue chips, when 1.5mm core tissue samples were punched from the original blocks to prepare tissue chips, there were no statistic difference among the positive rate by using the three methods in punching core samples, although the positive coincidence rate between the expression of ER and PR on conventional tissue section and tissue chip was on the rise with the increase of the number of core tissues punched from one original block. It could be concluded that tissue chip technique could substitute for conventional tissue section technique in immunohistochemistry, in situ hybridization（ISH） and fluorescence in situ hybridization（FISH）, and one core tissue punched from each original block could ensure the reliability and high-throughput characters of tissue microarray technique.