Evaluation Expression of miR-146a and miR-155 in Non-Small-Cell Lung Cancer Patients

BackgroundNon-small-cell lung cancer (NSCLC) is the major type of lung cancer. MicroRNAs (miRNAs) are novel markers and targets in cancer therapy and can act as both tumor suppressors and oncogenes and affect immune function. The aim of this study was to investigate the expression of miR146a and miR155 in linked to blood immune cell phenotypes and serum cytokines in NSCLC patients. MethodsThirty-three NSCLC patients and 30 healthy subjects were enrolled in this study. The allele frequencies of potential DNA polymorphisms were studied using polymerase chain reaction (PCR)-restriction fragment length polymorphism (PCR-RFLP) analysis in peripheral blood samples. Quantitative reverse transcription PCR (qRT-PCR) was used to measure the expression of miR-146a and miR-155 in peripheral blood mononuclear cells (PBMCs). Serum cytokine (IL-1 beta, IL-6, TNF-alpha, TGF-beta, IL-4, IFN-gamma) levels were determined by ELISA. The frequency of circulating CD3+CTLA-4+ and CD4+CD25+FOXP3+ (T regulatory cells/Treg) expression was measured by flow cytometry. ResultsmiR-146a was significantly downregulated in PBMC of NSCLC patients (P <= 0.001). Moreover, IL-6 and TGF-beta levels were elevated in NSCLC patients (P <= 0.001, P <= 0.018, respectively). CD3+ CTLA-4+ and Treg cells frequencies were higher in patients than in control subjects (P <= 0.0001, P <= 0.0001, respectively). There was a positive correlation between miR-155 and IL-1 beta levels (r=0.567, p <= 0.001) and a negative correlation between miR-146a and TGF-beta levels (r=-0.376, P <= 0.031) in NSCLC patients. No significant differences were found in the relative expression of miR-146a and miR-155, cytokine levels or immune cell numbers according to miR-146a and miR-155 (GG/GC/CC, TT/AT/AA) genotypes. However, there was a positive correlation between miR-146a and IL-1 beta levels (r=0.74, P <= 0.009) in GG subjects and a positive correlation between miR-146a expression and CD3+CTLA4+ cell frequency (r=0.79, P <= 0.01) in CC genotyped subjects. Conversely, a negative correlation between miR-146a expression and Treg cell frequency (r=-0.87, P <= 0.05) was observed with the GG genotype. A positive correlation between miR-155 and IL-1 beta expression (r=0.58, p <= 0.009) in the TT genotype and between miR-155 expression and CD3+CTLA-4 cell frequency (r=0.75, P <= 0.01) was observed in the AT genotype. ConclusionsThe current data suggest that the miR-146a expression in PBMC and serum TGF-beta and IL-1 beta levels may act as blood markers in NSCLC patients. Further study is needed to elucidate the link between immune cells and serum miR146 at early disease stages.

Automated summary

The study found that miR-146a expression was significantly downregulated in NSCLC patients, while IL-6 and TGF-beta levels were elevated. The frequencies of immune cells CD3+ CTLA-4+ and Treg cells were also higher in patients.