engleski

Flow and mass cytometry assessment of cervical precancerous lesions: preliminary study

Cervical cancer is one of the most common malignancies among female population despite existing prevention options. The development of cervical cancer is preceded by precancerous changes called squamous intraepithelial lesions (SIL) that usually develop over several years. SIL is divided into two stages: low-grade squamous intraepithelial lesions (LSIL) and high-grade squamous intraepithelial lesions (HSIL) depending on severity. Cervical cancer and SILs are associated with persistent high-risk human papillomavirus (HR HPV) infection that is sexually transmitted and responsible for 90-100% of cervical cancer cases among women. Although cervical cancer can be prevented by vaccination, cytological and molecular screening of sexually active women, lack of organized screening programs and/or vaccine hesitancy limit primary preventive measures in some populations. Secondary prevention measures including treatment of precancer come with associated risks and socioeconomic costs despite being highly successful. Due to the above, cervical cancer will remain a significant public health issue in the near future. In this study, gynecological clinical samples were collected with cytobrush and stored in NovaPrep liquid-based cytology (LBC) medium. Samples were classified as normal, LSIL and HSIL by an experienced cytologist. HPV genotyping by Multiplex PCR was performed on the isolated DNA. NovaPrep fixed HeLa cell line and clinical samples were assessed for the presence of several markers including p16, MCM2 and Ck18 by flow cytometry on FACSCalibur instrument. Preliminary data suggest that NovaPrep fixed HeLa cells can be reliably stained with the analyzed markers but protocols need to be further optimized for clinical samples. Further plans are to assess more biomarker candidates and include the information collected through a 2-year follow up period to find those associated with disease or virus persistence or progression.

Biomarkers ; Cervical cancer ; HPV

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