Identification and performance evaluation of house... - BV FAPESP
Advanced search
Start date
Betweenand
Related content
(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Identification and performance evaluation of housekeeping genes for microRNA expression normalization by reverse transcription-quantitative PCR using liquid-based cervical cytology samples

Full text
Author(s):
Causin, Rhafaela Lima [1] ; Pessoa-Pereira, Danielle [1] ; Borba Souza, Karen Cristina [1] ; Evangelista, Adriane Feijo [1] ; Vieira Reis, Rui Manuel [1] ; Tavares Guerreiro Fregnani, Jose Humberto [2] ; Chiquitelli Marques, Marcia Maria [1, 3]
Total Authors: 7
Affiliation:
[1] Barretos Canc Hosp, Mol Oncol Res Ctr, 1331 Rua Antenor Duarte Villela, BR-14784400 Barretos, SP - Brazil
[2] AC Camargo Canc Ctr, BR-01509010 Sao Paulo, SP - Brazil
[3] Barretos Sch Hlth Sci FACISB, BR-14785002 Barretos, SP - Brazil
Total Affiliations: 3
Document type: Journal article
Source: Oncology Letters; v. 18, n. 5, p. 4753-4761, NOV 2019.
Web of Science Citations: 0
Abstract

Screening for cervical cancer by cytology has been effective in reducing the worldwide incidence and mortality rates of this disease. However, a number of studies have demonstrated that the sensitivity of conventional cervical cytology may be too low for detection of cervical intraepithelial neoplasias (CIN). Therefore, it is important to incorporate more sensitive molecular diagnostic tests that could substantially improve the detection rates and accuracy for identifying CIN lesions. MicroRNAs (miRNAs) are a class of small non-coding RNAs with the potential to provide robust non-invasive cancer biomarkers for detecting CIN lesions in liquid-based cervical cytology (LBC) samples. At present, there is no consensus on which are the best housekeeping genes for miRNA normalization in LBC. The present study aimed to identify housekeeping genes with consistent and reproducible performance for normalization of reverse transcription-quantitative PCR (RT-qPCR) expression analysis of miRNA using LBC samples. The present study firstly selected six potential candidate housekeeping genes based on a systematic literature evaluation. Subsequently, the expression levels of microRNAs U6, RNU-44, RNU-47, RNU-48, RNU-49 and hsa-miR-16 were measured in 40 LBC samples using RT-qPCR. The stability of each potential housekeeping gene was assessed using the NormFinder algorithm. The results revealed that U6 and RNU-49 were the most stable genes among all candidates requiring fewer amplification cycles and smaller variation across the sample set. However, RNU-44, RNU-47, RNU-48 and hsa-miR-16 stability exceeded the recommended housekeeping value suitable for normalization. The findings revealed that U6 may be a reliable housekeeping gene for normalization of miRNA RT-qPCR expression analysis using LBC samples. (AU)

FAPESP's process: 16/15831-3 - Molecular signatures of microRNAs in precursor lesions of cervical cancer
Grantee:Rhafaela Lima Causin
Support Opportunities: Scholarships in Brazil - Master