Increased Cycling Cell Numbers and Stem Cell Associated Proteins as Potential Biomarkers for High Grade Human Papillomavirus+ve Pre-Neoplastic Cervical Disease

Abstract

High risk (oncogenic) human papillomavirus (HPV) infection causes cervical cancer. Infections are common but most clear naturally. Persistent infection can progress to cancer. Pre-neoplastic disease (cervical intraepithelial neoplasia/CIN) is classified by histology (CIN1-3) according to severity. Cervical abnormalities are screened for by cytology and/or detection of high risk HPV but both methods are imperfect for prediction of which women need treatment. There is a need to understand the host virus interactions that lead to different disease outcomes and to develop biomarker tests for accurate triage of infected women. As cancer is increasingly presumed to develop from proliferative, tumour initiating, cancer stem cells (CSCs), and as other oncogenic viruses induce stem cell associated gene expression, we evaluated whether presence of mRNA (detected by qRT-PCR) or proteins (detected by flow cytometry and antibody based proteomic microarray) from stem cell associated genes and/or increased cell proliferation (detected by flow cytometry) could be detected in well-characterised, routinely collected cervical samples from high risk HPV+ve women. Both cytology and histology results were available for most samples with moderate to high grade abnormality. We found that stem cell associated proteins including human chorionic gonadotropin, the oncogene TP63 and the transcription factor SOX2 were upregulated in samples from women with CIN3 and that the stem cell related, cell surface, protein podocalyxin was detectable on cells in samples from a subset of women with CIN3. SOX2, TP63 and human gonadotrophin mRNAs were upregulated in high grade disease. Immunohistochemistry showed that SOX2 and TP63 proteins clearly delineated tumour cells in invasive squamous cervical cancer. Samples from women with CIN3 showed increased proliferating cells. We believe that these markers may be of use to develop triage tests for women with high grade cervical abnormality to distinguish those who may progress to cancer from those who may be treated more conservatively.

Human papilloma virus: Apprehending the link with carcinogenesis and unveiling new research avenues (Review)

“…The use of cervical-vaginal fluid samples has demonstrated benefits as a biomarker source for lower female reproductive system pathologies, due to easy collection, with the prospective of utilizing self-tests. The following detection methods have been used, to date, for this sample examination: RT-qPCR for mRNA, flow cytometry and antibody-based proteomic microarray for stem cell-associated genes and flow cytometry for increased cell proliferation ( 121 ).…”

“…In high-grade disease samples, SOX2 , TP63 and human gonadotrophin mRNAs were upregulated. Immunohistochemistry experiments performed revealed that SOX2 and TP63 proteins visibly outlined tumor cells in invasive forms of squamous cervical cancer ( 121 ).…”

Expression of human papillomavirus L1 protein and NuMA1 at cervical intraepithelial neoplasia

“…Изменения экспрессии клеточных белков Ki-67 [21,24,25], PCNA [26,27], p53 [21,28], p16(INK4a) [21,24,25,29], Bcl-2 [28], OCT-4 [30], COX-2 [24], SOX2, р63 [31] оказались более информативны в качестве дифференциальных признаков цервикальной неоплазии разной степени тяжести [21,29]. Их корреляция с тяжестью цитологических и(или) гистологических поражений [32] может быть обусловлена механизмом, не связанным с вирусным поражением клетки [33].…”

Nitrative and oxidative DNA damage in infection-related carcinogenesis in relation to cancer stem cells

“…Stem cell associated proteins including human chorionic gonadotropin, the oncogene TP63 and the transcription factor SOX2 were upregulated in samples from women with CIN3 [55]. The stem cell related, cell surface protein podocalyxin was detectable on cells in samples from a subset of women with CIN3.…”

“…The stem cell related, cell surface protein podocalyxin was detectable on cells in samples from a subset of women with CIN3. SOX2 and TP63 proteins clearly delineated tumour cells in invasive squamous cervical cancer [55]. …”