Evaluation of RNA Markers for Early Detection of Cervical Neoplasia in Exfoliated Cervical Cells

Abstract

Numerous molecular biomarkers have been suggested for early detection of cervical cancer, but their usefulness in routinely collected exfoliated cells remains uncertain. We used quantitative reverse transcription-PCR to evaluate expression of 40 candidate genes as markers for high-grade cervical intraepithelial neoplasia (CIN) in exfoliated cervical cells collected at the time of colposcopy. Samples from the 93 women with CIN3 or cancer were compared with those from 186 women without disease matched (1:2) for age, race, and high-risk human papillomavirus status. Normalized threshold cycles (C t ) for each gene were analyzed by receiver operating characteristics to determine their diagnostic performance in a split sample validation approach. Six markers were confirmed by an area under the curve >0.6 in both sample sets: claudin 1 (0.75), minichromosome maintenance deficient 5 (0.71) and 7 (0.64), cell division cycle 6 homologue (0.71), antigen identified by monoclonal antibody Ki-67 (0.66), and SHC SH2-domain binding protein 1 (0.61). The sensitivity for individual markers was relatively low and a combination of five genes to a panel resulted in 60% sensitivity with 76% specificity, not positively increasing this performance. Although the results did not indicate superiority of RNA markers for cervical cancer screening, their performance in detecting disease in women referred for colposcopy suggests that the genes and pathways they highlight could be useful in alternative detection formats or in combination with other screening indicators. (Cancer Epidemiol Biomarkers Prev 2007;16(2):295 -301)

Reverse transcription-quantitative polymerase chain reaction: description of a RIN-based algorithm for accurate data normalization

“…Thus, any variation in CAB expression level would reflect variations in the efficiency of the RT and/or PCR steps. CAB showed a 1.5-fold variation range in our cell line cDNA samples, which is comparable to or even narrower than previously reported values for similar exogenous controls [ 6 , 19 , 20 ]. We conclude that in our samples and under our optimized RT-qPCR conditions, there was only a negligible effect of inhibitors on the RT and PCR efficiencies.…”

“…These inhibitors, which may include reagents used during RNA isolation, or co-purified biological components [ 16 , 17 ], can reduce the efficiency of both RT and PCR and generate errors in the quantification results. In this study, we used an exogenous CAB mRNA control [ 18 , 19 ] that was co-reverse-transcribed with each sample RNA and then amplified by qPCR. Thus, any variation in CAB expression level would reflect variations in the efficiency of the RT and/or PCR steps.…”

In SilicoAnalysis Identifies a Novel Role for Androgens in the Regulation of Human Endometrial Apoptosis

“…For example, SHCBP1 (SHC SH2 binding protein 1) has been cited as one of a panel of six markers for the diagnosis of early cervical cancer (32). PDGFRL , a gene that encodes a secreted protein with significant sequence homology to the ligand-binding domain of platelet-derived growth factor receptor-β, has been proposed as a tumor suppressor (33).…”

Potential role of cancer stem cells as biomarkers and therapeutic targets in cervical cancer

“…Radiological techniques and circulating levels of tumor‐specific antigens are useful in assessing traditionally known biomarkers. Technologies like mass spectrometry, microarrays, high‐throughput DNA sequencing, complete human genome sequence, and the information on potential cancer biomarkers are useful to know the DNA, RNA, and protein expressions . Advances in imaging technologies will give way to noninvasively monitor molecular biomarkers (eg, those responding to therapy) in cancer patients …”

“…Still, there are major challenges in the CSC field (Table 3) 37,39,40,59 Advances in imaging technologies will give way to noninvasively monitor molecular biomarkers (eg, those responding to therapy) in cancer patients. 7,90 The current understanding of the concepts of cervical CSC biomarkers and their evaluation in clinical scenario is suggestive that Abbreviations: ALDH1, aldehyde dehydrogenase 1; CCSCs, cervical cancer stem cells; CD133, cluster of differentiation 133(Prominin-1); CD24, cluster of differentiation 24 (heat stable antigen-HSA); CIN, cervical intraepithelial neoplasia; CK17, cytokeratin 17; OCT4, octamer-binding transcription factor 4; PIWIL1, piwi-like RNA-mediated gene silencing 1; PSCA, prostate stem cell antigen; SOX2, sex-determining region-Y-box 2.…”