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

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.