免疫组织化学检测在胸水液基细胞学中的可行性研究

Feasibility study of immunohistochemistry for liquid-based cytology in pleural exfoliation examination

背景与目的:液基细胞学(liquid-based cell test,LCT)是胸水脱落细胞诊断的常规技术,鉴别诊断时需用细胞学包块行免疫组织化学检查。运用LCT直接行免疫组织化学检测,探讨其方法的可行性及临床意义。方法:回顾性分析复旦大学附属华东医院病理科2016—2017年间收集的96例胸水脱落细胞学检查标本,运用PrepStain自动细胞学制片机薄层细胞学H-E染色片及白片若干张,抗原修复后进行甲状腺转录因子1(thyroid transcription factor 1,TTF1)、P63、WT1、雌激素受体(estrogen receptor,ER)、孕激素受体(progesterone receptor,PR)、人表皮生长因子受体2(human epidermal growth factor receptor 2,HER2)、CK7、CK20、CDX2、CA19-9、P53、PAX2及TG等免疫组织化学染色,观察染色结果,结合临床病史综合分析。结果:46例胸水H-E染色片内找见癌细胞,通过免疫组织化学染色并结合临床病史,发现TTF-1阳性的肺腺癌19例(占恶性组59.3%),P63阳性的鳞状细胞癌3例,ER/PR/HER2染色阳性的转移性乳腺癌3例,WT1/P53/ER/PAX2染色阳性的转移性浆液性癌2例,CDX2染色阳性的转移性结肠癌2例,TTF1/TG染色阳性的转移性甲状腺滤泡癌1例,CA19-9阳性的胰腺癌2例。经TTF1(-)/P63(-)/WT1(+)染色明确反应性间皮细胞43例。仍有7例经免疫组织化学染色无法区分反应性间皮细胞和癌细胞、14例低分化癌细胞无法判断来源,原因是所选的特异性抗原均不表达、异型细胞量少在免疫组织化学片内消失或肿瘤细胞脱片。结论:相比较细胞学包块检查过程,薄层细胞学制片方法简单省时,经适当抗原修复,免疫组织化学染色结果可靠,细胞核和细胞膜的抗原表达易于观察。免疫组织化学运用于胸水薄层细胞学制片检查可提高诊断的准确率和特异度,为临床治疗提供可靠依据。

Background and purpose:Liquid-based cell test(LCT)is a routine method in pleural exfoliation examination,while cell block is required to perform immunohistochemistry for differential diagnosis.In this study,we used liquid-based cytology directly for immunohistochemical staining and discussed the feasibility of the method and its clinical significance.Methods:A total of 96 cases of pleural effusion were collected from 2016 to 2017 in Department of Pathology,Huadong Hospital Affiliated to Fudan University to prepare a H-E cytologic staining and white slices by PREPSTAIN thin liquid-based cell test machine.Immunohistochemical staining of cytologic white slices for thyroid transcription factor 1(TTF1),P63,WT1,estrogen receptor(ER),progesterone receptor(PR),human epidermal growth factor receptor 2(HER2),CK7,CK20,CDX2,CA19-9,P53,PAX2 and TG was performed after antigen repairing,and staining results were interpreted and analyzed with clinical history comprehensively.Results:Carcinoma was found in 46 cases,including 19 cases of lung adenocarcinoma confirmed by positive TTF1 staining(accounted for 59.3%of all carcinomas),3 cases of squamous cell carcinoma by positive P63,3 cases of metastatic breast carcinoma by positive ER/PR/HER2,2 cases of metastatic ovary serous carcinoma by positive WT1/P53/ER/PAX2,2 cases of metastatic colon adenocarcinoma by positive CDX2,1 case of metastatic thyroid follicular carcinoma by positive TTF1/TG and 2 cases of metastatic pancreatic carcinoma by positive CA19-9.Forty-three cases of reactive mesothelial cells were confirmed by TTF1(-)/P63(-)/WT1(+)staining.There were still 7 cases hard to differentiate reactive from malignant cells after immunohistochemical staining,and 14 casesof poorly differentiated carcinoma in which the organ source cannot be determined due to no specific antigen expression,limitedatypical cells and detachment in immunohistochemical procedure.Conclusion:Liquid-based cytology procedure is simpler and moretime-saving than the method of cell block.The immunohistochemical result of cytology is reliable after proper antigen repairing andinterpretable for the nuclear and membrane expression.Immunohistochemical staining performed using liquid-based cytology ofpleural exfoliation may improve the accuracy and specificity of diagnosis and provide reliable basis for clinical treatment.