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

Abstract

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.

Keywords

• Cervical cancer
• Aneuploid TCs and TECs
• iFISH tumor tissue biopsy
• Chromosome 8
• p16 and Ki67
• HPV16/18