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.