Over Expressed TKTL1, CIP-2A, and B-MYB Proteins in Uterine Cervix Epithelium Scrapings as Potential Risk Predictive Biomarkers in HR-HPV-Infected LSIL/ASCUS Patients

Abstract

High oncogenic risk human papillomaviruses (HR-HPVs) promote cervical carcinoma development, the fourth most common feminine cancer. A slow oncodevelopmental phase—defined histopathologically as Cervical Intraepithelial Neoplasia (CIN) grades 1–3, or cytologically as Low- or High-grade Squamous Intraepithelial Lesions (LSIL or HSIL)—precedes the malignancy. Cervical carcinoma screenings through HR-HPV genotyping and Pap smears are regularly performed in Western countries. Faulty cytology screening or genotyping or patients' non-compliance with follow-ups can let slip an oncoprogression diagnosis. Novel biomarker tests flanking HR-HPV genotyping and cytology could objectively predict the risk of disease progression thus helping triage LSIL/ASCUS patients. Here, anonymized leftovers of fresh cervical epithelium scrapings from twice (LSIL/ASCUS and HR-HPV DNA)-positive and twice (Pap smear- and HR-HPV DNA)-negative (control) patients in a proteome-preserving solution served to assess the biomarker worth of three cervical carcinoma-related proteins, i.e., B-MYB (or MYBL2), Cancerous Inhibitor of PP2A (CIP-2a), and transketolase-like1 (TKTL1). Leftovers anonymity was strictly kept and storage at −80°C, protein extraction, immunoblotting, and band densitometry were blindly performed. Only after tests completion, the anonymous yet code-corresponding HR-HPV-genotyping and cytology data allowed to assign each sample to the twice-positive or twice-negative group. Descriptive statistics showed that the three proteins levels significantly increased in the twice-positive vs. twice-negative scrapings. Diagnostic ROC curve analysis identified each protein's Optimal Decision Threshold (OTD) showing that TKTL1 and CIP-2a are stronger risk predictive biomarkers (Sensitivity, 0.91–0.93; Specificity, 0.77–0.83) than B-MYB. Logistic Regression coupled with Likelihood-Ratio Tests confirmed that a highly significant relation links increasing TKTL1/CIP-2a/B-MYB protein levels in twice-positive cervical scrapings to the risk of HR-HPV-driven oncoprogression. Finally, a 3 year clinical follow-up showed that 13 patients (50% of total) of the twice-positive group with biomarker values over OTDs compliantly underwent scheduled colposcopy and biopsy. Of these, 11 (i.e., 84.7%) received a positive histological diagnosis, i.e., CIN1 ( n = 5; 38.5%) or CIN2/CIN2+ ( n = 6; 46,2%). Therefore, TKTL1/CIP-2a/B-MYB protein levels could objectively predict oncoprogression risk in twice (HR-HPV- and Pap smear)-positive women. Further studies will assess the translatability of these findings into clinical settings.

Use of AKR1C1 and TKTL1 in the Diagnosis of Low-grade Squamous Intraepithelial Lesions from Mexican Women

“…In the same way as AKR1C1, an increase in the expression of TKTL1 was observed both in LSIL and HPV-positive samples. Recent studies have shown a high expression of this protein in patients diagnosed with atypical squamous cells of undetermined significance and HPV (41), which confirms our results in LSIL histological samples.…”

“…Therefore, the biomarkers proposed in this work can be used for the diagnosis of LSIL, where the specificity of other markers is low. In addition, the sensitivity and specificity of AKR1C1 with TKTL1 in HSIL and invasive CC should be evaluated, since the combination of TKTL1 and cancerous inhibitor of protein phosphatase 2 (CIP-2A) as biomarkers predictive of risk had a sensitivity of 0.91-0.93 and specificity of 0.77-0.83, with high expression in double-positive patients with high-risk HPV-DNA genotype infection and diagnosis of atypical glandular cells of undermined significance (41).…”

TKTL1 participated in malignant progression of cervical cancer cells via regulating AKT signal mediated PFKFB3 and thus regulating glycolysis

“…In addition, the increased expression of TKTL1 has been proved to be a diagnostic marker in low-grade CC [ 15 ]. At the same time, study has confirmed that TKTL1 expression is increased in CC and can be used as a diagnostic marker for CC [ 16 ]. Since TKTL1 has a cancer-promoting biological activity, this paper aims to explore whether TKTL1 can participate in the malignant process of CC cells through the glucose metabolism pathway.…”

Changes in the Proteome in the Development of Chronic Human Papillomavirus Infection—A Prospective Study in HIV Positive and HIV Negative Rwandan Women

“…Proteomics has been used for identifying biomarkers correlating with the presence of premalignant lesions and cancer of the uterine cervix [ 7 , 8 ]. Biomarkers may not only be identified directly in the uterine cervix but also in the cervicovaginal fluid [ 9 , 10 ], which makes self-sampling possible for screening.…”