5 research outputs found
microRNA Profile Associated with Positive Lymph Node Metastasis in Early-Stage Cervical Cancer
Lymph node metastasis (LNM) is an important prognostic factor in cervical cancer (CC). In early stages, the risk of LNM is approximately 3.7 to 21.7%, and the 5-year overall survival decreases from 80% to 53% when metastatic disease is identified in the lymph nodes. Few reports have analyzed the relationship between miRNA expression and the presence of LNM. The aim of this study was to identify a subset of miRNAs related to LNM in early-stage CC patients. Formalin-fixed paraffin-embedded tissue blocks were collected from patients with early-stage CC treated by radical hysterectomy with lymphadenectomy. We analyzed samples from two groups of patients—one group with LNM and the other without LNM. Global miRNA expression was identified by microarray analysis, and cluster analysis was used to determine a subset of miRNAs associated with LNM. Microarray expression profiling identified a subset of 36 differentially expressed miRNAs in the two groups (fold change (FC) ≥ 1.5 and p < 0.01). We validated the expression of seven miRNAs; miR-487b, miR-29b-2-5p, and miR-195 were underexpressed, and miR-92b-5p, miR-483-5p, miR-4534, and miR-548ac were overexpressed according to the microarray experiments. This signature exhibited prognostic value for identifying early-stage CC patients with LNM. These findings may help detect LNM that cannot be observed in imaging studies
Comparison of Automated and Manual DNA Isolation Methods of Liquid-Based Cytology Samples
Liquid-based cytology (LBC) has been used as a diagnostic tool for cervical cancer for years and is now being
adopted for other gynecological cancers. LBC represents an important challenge to ensure that the process yields
representative biospecimens for quality control (QC) of diagnostic procedures. In this study, we compare QC
parameters (integrity, yield and purity, and polymerase chain reaction [PCR] amplification) of DNA isolated from
LBC (N= 296) using two different nucleic acid isolation methods, manual (n = 233) or automated (n = 63). We also
evaluated two different types of cytological brushes for sampling from the cervix. Our results suggest that manual
isolation (yield 22.81 – 1.92 mg) resulted in increased DNA recovery when compared with automated isolation
(yield 9.96 – 1.11 mg) from LBC samples, with a p-value of <0.0003. We estimated that 98% (53/54) of the samples
preserved the integrity of DNA and were suitable for standard molecular biology analyses. The b-globin gene was
amplified in 100% (296/296) of the DNA samples by endpoint PCR. We found no significant difference between
the performance of the cytological brushes ( p value of <0.6711) in a general overview. However, when looking
at the results from using each brush individually, the manual isolation method was statistically superior to the
automated method. Our work illustrates the impact of good QC of preanalytic conditions, which will be
important for the application of LBC for developing early detection methods for gynecological cancers