Abstract
BackgroundCirculating cell-free microRNAs have been identified as potential cancer biomarkers. However, the existence and the potential application of cell-free miRNAs in effusion samples are still uncertain. In order to explore the potential role of cell-free miRNA in malignant effusions, we selected 22 miRNAs differentially expressed in the serum of lung cancer patients and studied their expression levels in body cavity effusion samples.MethodsWe measured the expression of 22 miRNAs using qRT-PCR in two samples, which were pooled with 18 malignant and 12 benign effusions, respectively. After discarding 9 lowly expressed miRNAs, a panel of 13 miRNAs were measured in 29 samples (benign n = 11, malignant n = 18). We also carried out a WST-8 test to evaluate the docetaxel sensitivity of tumor cells directly isolated from 15 malignant effusions.ResultsWe compared the miRNA expression levels between benign and malignant effusions using a Mann-Whitney U test and found miR-24, miR-26a and miR-30d were expressed differently between the two groups (P = 0.006, 0.021 and 0.011, respectively). Cells isolated from effusions rich in cell-free miR-152 were more sensitive to docetaxel (r = 0.60, P = 0.016).ConclusionsCollectively, our study demonstrated that cell-free miRNAs in the supernatant of effusions may aid in the diagnosis of malignancy and predict chemosensitivity to docetaxel.
Highlights
Circulating cell-free microRNAs have been identified as potential cancer biomarkers
The stability of miRNAs in the effusion samples We firstly tested the stability of cell-free miRNAs in the effusion samples kept at room temperature for different time periods, or treated with multiple freezing and thawing, and digestion with RNase or DNase. bactin mRNA transcripts and 18S RNA representing large molecular RNAs were analyzed in this study. Quantitative RT-PCR (qRT-PCR) was carried out to quantify the RNAs of three samples with various treatments
When effusion samples were left at room temperature for 3 h or 24 h, cell-free RNAs began to degrade at 3 h. b-actin mRNA degraded quickly and significantly, while 18S RNA and the two miRNAs were more stable, with more than 20% remaining after 24 h (Figure 1B)
Summary
Circulating cell-free microRNAs have been identified as potential cancer biomarkers. The existence and the potential application of cell-free miRNAs in effusion samples are still uncertain. In order to explore the potential role of cell-free miRNA in malignant effusions, we selected 22 miRNAs differentially expressed in the serum of lung cancer patients and studied their expression levels in body cavity effusion samples. Body cavity effusion is a clinical common manifestation and may cause dilemmas in treatment. Diagnosis of effusions is of particular importance for cancer patients and a co-existing malignant effusion implies an advanced stage of tumor and intricate clinical management. Diagnosis of malignant effusions mainly relies on cytological analysis. Alternative diagnostic methods are still in need to assist in the diagnosis of effusions.
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