细胞DNA倍体分析评估宫颈上皮内瘤变

目的评价通过细胞DNA倍体分析诊断宫颈上皮内瘤变（CIN）的效果。方法对87例宫颈癌普查妇女用宫颈刷取材、液基薄层制片,由细胞学医师对巴氏染色片做常规细胞诊断,应用全自动DNA倍体分析仪对DNA染色片进行自动扫描诊断。对活检样本做出病理诊断,并应用免疫组化ABC法测定宫颈鳞状上皮细胞核内Ki67蛋白的表达。结果87例受检者中,常规细胞学诊断为正常30例,非典型鳞状细胞（ASCUS）20例,低度鳞状上皮内病变（LSIL）30例,高度鳞状上皮内病变（HSIL）7例;无异倍体细胞者26例,有异倍体细胞者61例,其中Ⅰ级17例,Ⅱ级21例,Ⅲ级23例。20例ASCUS者中,有7例无异倍体细胞,未发现一例CIN2以上级别的病理改变,Ki67阳性细胞分布在鳞状上皮下方37．1％范围内;13例有异倍体细胞者中,11例有CIN2以上病理改变,Ki67免疫阳性细胞分布于上皮层下46．2％范围内。30例LSIL者中,有7例未发现异倍体细胞,其中3例有CIN2以上病理改变,Ki67阳性细胞只分布在鳞状上皮下方37．1％范围内;23例伴有DNA异倍体细胞者中,22例发现CIN2以上病理改变,Ki67阳性细胞增至鳞状上皮层中上方62．6％范围内,与7例未发现异倍体细胞者差异有统计学意义（P〈0．01）。7例HSIL病例中均有DNA异倍体细胞出现。以≥3个DNA异倍体细胞作为评估CIN2及以上病理改变的标准,其敏感性为72．7％,特异性为87．5％,阳性预测值为90．9％,阴性预测值为65．1％,均比以常规细胞学诊断为LSIL及以上级别作为评估CIN2及以上病理改变的标准（分别为58．2％、84．4％、86．5％和54．0％）要高。结论DNA倍体分析可诊断宫颈CIN病变,并预测其发展。

Assessment of cervical intraepithelial neoplasia (CIN) lesions by DNA image cytometry

OBJECTIVE: To compare the value of conventional cytology and DNA image cytometry (DNA-ICM) assisted cytology in detection and prognostic assessment of cervical CIN lesions. METHODS: 87 women were enrolled in this study. Cervical samples were collected employing cervix brushes which were then washed in Sedfix. After preparing single cell suspensions by mechanical procedure, cell monolayers were prepared by cyto-spinning the cells onto microscope slides. Two slides were prepared from each case: one slide was stained by Papanicolou staining for conventional cytology, another was stained by Feulgen-Thionin method for measurements of the amount of DNA in the cell nuclei using an automated DNA imaging cytometer. Biopsies from the cervical lesions were also taken for histopathology and Ki-67 immunohistochemistry. RESULTS: Of the total of 20 ASCUS cases called by conventional cytology, no CIN, nor greater lesions were found. Among the 20 cases, 7 cases did not show any cells with DNA amount greater than 5c, while CIN2 lesions were found in 11 of other 13 cases that had some aneuploid cells with DNA amount greater than 5c. Of 30 LSIL cases called by conventional cytology, CIN2 lesions were detected in 3 out of 7 cases that did not contain any aneuploid cells with DNA greater than 5c, but in 22 out of the other 23 cases that contained aneuploid cells with DNA amount greater than > 5c. Of the remaining 7 cases called HSIL by conventional cytology, all case contained aneuploid cells containing DNA greater than 5c. If cytology was used to refer all cases of LSIL and HSIL to colposcopy procedure to detect potential CIN2 or greater lesions, the sensitivity, specificity, positive predictive value and negative predictive value were 58.2%, 84.4%, 86.5% and 54.0%, respectively. If DNA-ICM were used and all cases having 3 or more cells with a DNA amount greater than 5c were assessed to be referred to pathology to detect potential CIN2 or greater lesions, the sensitivity, specificity, positive predictive value and negative predictive were 72.7% , 87.5%, 90.9% and 65.1%, respectively. We also compared Ki67 positive cells in these samples and found that DNA-ICM results were comparable to this biomarker method. CONCLUSION: The study demonstrated that DNA-ICM approach can be successfully used to detect significant (i.e. CIN2 or greater) lesions, and also provide a prognostic assessment of CIN lesions.