HPVE6/E7,hTERT, andKi67mRNA RT-qPCR Assay for Detecting High-Grade Cervical Lesion with Microscope Slides

Abstract

After breast and colon cancer, cervical cancer is the third most common cancer of women worldwide. Since human papillomavirus (HPV) infection is known to be the predominant cause of cervical cancer, molecular HPV screening is currently used along with cytological and histological examination methods for precancer diagnosis. Nevertheless, the sensitivity of the current HPV test is less than 80%; thus, many cervical cancer cases are not able to be diagnosed by HPV screening alone, and likewise, patients with cervical cancer are often determined to be HPV-negative by the current screening methods. Therefore, human telomerase reverse transcriptase (hTERT) and Ki67 previously identified as cancer markers were attempted. And cervical exfoliated cells of high-grade squamous intraepithelial lesion (HSIL), the most severe precancerous lesion of cancer, were used in the study. However, it takes a long time to collect enough specimens to conduct statistical analysis. Therefore, in the present study, microscope slides, cervical exfoliated cells on glass slides, were attempted. The results of the analysis demonstrated that hTERT and Ki67 expression levels were useful in distinguishing between cancerous and normal specimens, exhibiting a higher sensitivity and specificity than conventional HPV E6/E7 testing. And the study suggests clinical slide cell samples could be effectively used in the context of retrospective studies to identify novel biomarkers.

Screening and Identification of New Biomarkers for the Control of Cervical Cancer

“…E6 inhibits p53 to prevent apoptosis, while E7 is the primary transforming protein and inhibits pRB to regulate cell cycle stop [51]. Telomeres are specialised structures, located at the ends of chromosomes, known to be essential for the stability of the genome [51,52]. Telomere dysfunction and telomerase activation have previously been implicated in the progression of human cancer [51,53].…”

Potential Biomarkers for Cervical Cancer Screening and Treatment

“…E6 and E7 are primary HPV oncoproteins with multiple cell targets, including p53, and the pRB. E6 inhibits p53 to prevent apoptosis, while E7 is the primary transforming protein and inhibits pRB to regulate cell cycle stop [76]. Telomeres are specialised structures, located at the ends of chromosomes, known to be essential for the stability of the genome [76,77].…”

“…E6 inhibits p53 to prevent apoptosis, while E7 is the primary transforming protein and inhibits pRB to regulate cell cycle stop [76]. Telomeres are specialised structures, located at the ends of chromosomes, known to be essential for the stability of the genome [76,77]. Telomere dysfunction and telomerase activation have previously implicated in the progression of human cancer [76,78].…”

“…Telomeres are specialised structures, located at the ends of chromosomes, known to be essential for the stability of the genome [76,77]. Telomere dysfunction and telomerase activation have previously implicated in the progression of human cancer [76,78]. The expression of hTERT is known to be the rate-limiting factor for human telomerase activity and a more sensitive indicator of telomerase function and action than the expression levels of other telomerase subunits that are expressed constitutively in normal and cancerous cells [76].…”

“…Telomere dysfunction and telomerase activation have previously implicated in the progression of human cancer [76,78]. The expression of hTERT is known to be the rate-limiting factor for human telomerase activity and a more sensitive indicator of telomerase function and action than the expression levels of other telomerase subunits that are expressed constitutively in normal and cancerous cells [76]. Ki67 is a nuclear antigen expressed during all active phases of the cell cycle (G1, S, G2 and M) and the level of expression can demonstrate to determine the state of proliferation and predict tumour development [76].…”