In situ phenotypic and karyotypic co-detection of aneuploid TCs and TECs in cytological specimens with abnormal cervical screening results.

Abstract

Background: To distinguish and co-detect aneuploid CD31^- tumor cells (TCs) and CD31⁺ tumor endothelial cells (TECs) may have significant diagnostic values for cervical cancer screening. However, there are very few relevant studies. In the present study, a novel "immunofluorescence staining integrated with fluorescence in situ hybridization (iFISH)" tumor tissue biopsy platform was applied to comprehensively investigate the clinical utilities of aneuploid TCs and TECs in all-stage cervical lesion smear specimens.

Methods: A total of 196 patients were enrolled in this study. Immunofluorescence staining of p16 and Ki67 combined with FISH was applied to quantitatively co-detect and characterize subcategorized aneuploid CD31^- TCs and CD31⁺ TECs in cervical cytological specimens. The Kruskal‒Wallis H test was used to compare the distributions of aneuploid TCs and TECs among all stages of cervical lesions and among the different high-risk HPV types (HPV16/18 and non-HPV16/18). The diagnostic value of detecting aneuploid TCs and TECs for high-grade squamous intraepithelial lesions (HSIL⁺) was investigated via receiver operating characteristic curve analysis.

Results: The number of total aneuploid CD31^- TCs and their p16⁺ and/or Ki67⁺ (p16/Ki67⁺) subtypes increased markedly with the severity of cervical lesions, although a similar trend was not observed for aneuploid CD31⁺ TECs. The increase in aneuploid TCs resulted from HPV16/18 infection was mainly concentrated in low-grade squamous intraepithelial lesion(LSIL), whereas the increase caused by non-HPV16/18 infection was mainly concentrated in HSIL. To identify HSIL⁺, the area under the curve (AUC) of tetraploid TCs was the largest (0.739), followed by multiploid (≥ pentaploid) TCs (0.724) and triploid TCs (0.699). For the combined subtypes, the AUC of ≥ tetraploid TCs was 0.745, and their unique diagnostic value was clinically reflected by their high specificity.

Conclusion: The quantity of CD31^- aneuploid TCs was associated with the severity of cervical lesions. In HPV16/18 positive patients, aneuploid CD31^- TCs were significantly increased in the LSIL. Moreover, aneuploid CD31^- TCs exhibited remarkable specificity for detecting HSIL⁺. Further studies are required to expand the potential clinical utility of detecting CD31^- aneuploid TCs.