Background/Objectives: Cervical Pap smears are routinely used to detect cellular abnormalities as a cervical cancer screening tool and to assess the presence of HPV for risk stratification of the disease. Here, we aimed to extend the applications of this sampling procedure by combining it with multicolor flow cytometry to characterize cell populations across cervical cancer disease stages.Methods: Cervical Pap smears from 30 patients with various disease stages ranging from normal to intraepithelial neoplasia up to treated cancers were analyzed as biofluids using multicolor flow cytometry. Individual samples were evaluated, and statistical analyses were performed over all sample stages. Cancer cell lines (CaSki, SiHa, HeLa, A549, U2OS) were examined as tumor cell controls.Results: Cervical biofluids were subdivided into cell populations according to their scattering properties and the expression of specific biomarkers: EpCAM and cytokeratin 8 for epithelial cells from tumors as well as healthy ectocervical and endocervical regions, and CD45 for immune cells. Discrimination of tumor cells was facilitated with cancer cell lines. Statistical analysis revealed that the composition of cell populations differs among disease stages, whereas treated cancer samples were consistently associated with a reduction in squamous epithelial cells and an increase in immune cells compared to normal samples.Conclusions: Herein, we identified the major cell populations in cervical biofluid samples and demonstrated that this method can detect changes in the cellular composition across different disease stages. This approach could be further exploited in cancer research and potentially serve as a companion diagnostic tool in tumor development, progression and during treatment.
背景/目的：宫颈巴氏涂片作为宫颈癌筛查工具，常规用于检测细胞异常并评估HPV感染情况以进行疾病风险分层。本研究旨在扩展该采样方法的应用范围，将其与多色流式细胞术相结合，以表征宫颈癌不同疾病阶段的细胞群特征。方法：收集30例涵盖从正常到上皮内瘤变直至治疗后癌症等不同疾病阶段的宫颈巴氏涂片样本，作为生物流体进行多色流式细胞术分析。对个体样本进行评估，并对所有疾病阶段样本进行统计分析。以癌细胞系（CaSki、SiHa、HeLa、A549、U2OS）作为肿瘤细胞对照。结果：根据散射特性及特异性生物标志物表达，将宫颈生物流体划分为不同细胞群：采用EpCAM和细胞角蛋白8标记来源于肿瘤组织、健康宫颈外口及宫颈管区域的上皮细胞，CD45标记免疫细胞。通过癌细胞系辅助实现了肿瘤细胞的鉴别。统计分析显示，不同疾病阶段的细胞群组成存在差异，其中治疗后癌症样本与正常样本相比，持续表现为鳞状上皮细胞减少和免疫细胞增加。结论：本研究明确了宫颈生物流体样本中的主要细胞群，并证明该方法能够检测不同疾病阶段细胞组成的变化。该技术可进一步应用于癌症研究，并有望作为肿瘤发生、发展及治疗过程中的辅助诊断工具。
肿瘤(癌症)患者之家