Use of hTERT and HPV E6/E7 mRNA RT-qPCR TaqMan Assays in Combination for Diagnosing High-Grade Cervical Lesions and Malignant Tumors

Abstract

Objectives: Human papillomavirus (HPV) is a major cause of cervical cancer, which is the second most common cancer in women. HPV E6 initiates degradation of cellular tumor suppressor protein p53, induces human telomerase reverse transcriptase (hTERT) activity, and then leads to progressive cervical carcinogenesis. Methods:Results: The positivity for the HPV E6/E7 messenger RNA (mRNA) assay was 94. 4%, 95.2%, 82.4%, 46.5%, 25.0%, and 1.1% in squamous cell carcinomas,, atypical squamous cellscannot exclude HSIL, low-grade squamous intraepithelial lesions, atypical squamous cells of undetermined significance, and normal cytology samples, respectively. Five cervical intraepithelial neoplasia grade 2+ samples were not detected by the HPV E6/E7 mRNA assay, but they exhibited positive signals in the hTERT mRNA assay. Notably, the hTERT mRNA expression level was increased in high-grade cervical lesions but was very low in all 288 normal samples. Conclusions:These data suggest that the combination of HPV E6/E7 and hTERT mRNA expression levels could be used in a complementary manner in diagnosing high-grade cervical lesions and malignant tumors and might be useful as a predictive marker in monitoring low-grade cervical lesions.Cervical cancer, which develops from cervical intraepithelial neoplasia (CIN), is the second most common malignancy in women worldwide and is a major cause of morbidity and mortality. 1 While the overall incidence of cervical cancer is declining, 493,000 new cases and 274,000 deaths occur each year. 2 Human papillomavirus (HPV) is the most common sexually transmitted pathogen among women and men, and it has been estimated that 70% of sexually active women will acquire an HPV infection at some point during their lifetime. HPVs are small double-stranded DNA viruses that infect basal cells and replicate within the nucleus of squamous epithelial cells. 3 Cervical cancer is caused by persistent infection with oncogenic HPV genotypes that belong to a few phylogenetically related high-risk (HR) species. Some HPV genotypes are able to infect cutaneous tissues, resulting in the formation of warts on the hands or feet, while others infect the oral mucosa. HPV genotypes that infect the genital epithelia are well characterized and can be grouped as HR species because of their association with cervical cancer. As a result, testing for oncogenic HPV infection in cervical lesions could serve as an accurate means of identifying women who are at risk for developing cervical cancer. HR HPV genotypes, including 16, 18, 33, 45, 52, 58, 39, 51, 56, and 59, are detected in more than 90% of cervical cancer cases. 4 The HR HPV genotypes express two major transforming genes that code for the E6 and E7 proteins, which are required for the immortalization of human primary genital keratinocytes. 5 These two oncogenes are uniformly retained and highly expressed in cervical cancer cells, and their continuous expression is required for retention of

Correlation of hTERT Expression with Cervical Cytological Abnormalities and Human Papillomavirus Infection

“…Significantly increased levels of the hTERT in high-grade and low-grade lesions compared to the normal samples suggest that hTERT can be used as an early diagnostic and prognostic marker. Our results are comparable to previous studies in which hTERT expression relates to the severity of the disease [30][31][32]. [33,34].…”

Prognostic role of vascular endothelial growth factor in cervical cancer: a meta-analysis

“…In order to improve survival outcome of cervical cancer, several prognostic biomarkers have been identified. Human-papilloma virus (HPV) related biomarkers have been proven as effective biomarkers in the diagnosis and prevention of cervical cancer [2, 3]. However, the researchers are still screening new markers which are highly associated with cervical cancer progression and prognosis.…”

Association Between Type-specific HPV Infections and hTERT DNA Methylation in Patients with Invasive Cervical Cancer

“…They proposed that a combination of the hTERT assay (showing high SP and PPV) with another test showing high SE and high NPV (as the HPV testing), could be an ideal screening tool for detection of CIN lesions (34). Recently, analyses of hTERT methylation or hTERT expression in combination with other biomarkers have been developed to improve the specificity in the detection of HSIL showing favourable results (35,(38)(39)(40). Future clinical studies should evaluate the use of hTERT methylation in combination with HPV species or HPV specific types, which could improve the detection of HSIL lesions.…”

“…Infections with high-risk human papillomavirus (hrHPV) types are detected in virtually all cervical carcinomas (2,3). At least 13 genotypes of the alpha genus (HPV types 16,18,31,33,35,39,45, 51, 52, 56, 58, 59, and 68) have been associated with the risk to develop cervical cancer and are defined as "carcinogenic" viral types (4)(5)(6). HPV16 and 18 are the most prevalent genotypes found in squamous cell carcinoma across the world (7).…”