Background: Hypoxia is a hallmark of cancer and is associated with poor prognosis. However, the molecular mechanism by which hypoxia promotes tumor development is still unclear. MicroRNAs dysregulation has been shown to play an important role in the tumor and the tumor micro. Here, we investigated the role of miR-495 and miR-5688 in lung cancer of human non-small cell (NSCLC) and their underlying mechanisms.
Methods: The expression level of miR-495 and miR-5688 in human NSCLC tissue specimens was measured by quantitative real-time polymerase chain reaction (qRT-PCR). Deferoxamine (DFO) is used to determine whether the regulation of miR-495 and miR-5688 under hypoxia depends on the hypoxia-inducible factor 1-alpha (HIF-1α).
Furthermore, the function of miR-495 and miR-5688 in tumor progression was evaluated using colony formation, 3- (4,5-dimethylthiazol-2-yl) -5- (3-carboxymethoxyphenyl) -2- (4-sulfophenyl) -2H -tetrazolium (MTS), wound healing, transwell test, and the xenograft model. Two algorithms, PicTAR and Targetscan, used to predict the two miRNAs target gene is, and dual-luciferase reporter assay was performed to confirm the target. Tests are two-tailed paired t, Pearson correlation analysis and Fisher probability test conducted for statistical analysis.
Results: The two miRNAs, miR-495 and miR-5688, was found to participate in the development of NSCLC under hypoxia. They were down-regulated in NSCLC tissue compared to normal tissue. We determined that hypoxia causes down-regulation of miR-495 and miR-5688 in NSCLC cells, independent of HIF-1α and energy metabolism of cells. In addition, miR-495 and miR-5688 suppressed cell proliferation, migration and invasion in vitro
miR-495 and miR-5688 are down-regulated in non-small cell lung cancer under hypoxia to maintain interleukin-11 expression
. The NSCLC xenograft models indicate that miR-495 and miR-5688 inhibits tumor formation in vivo. Interestingly, we found that miR-495 and miR-5688 have the same target, interleukin-11 (IL-11). Human recombinant IL-11 is counteracted the effects of miR-495 and miR-5688 in NSCLC cells, suggesting that miR-495 and miR-5688 held their tumor suppressor role by pressing IL-11 expression.
Conclusion: We found hypoxia down-regulated the expression level of miR-495 and miR-5688 in NSCLC to increase IL-11 expression and tumor progression, suggesting that miR-495 / miR-5688 / IL-11 axis can serve as a therapeutic target and a potential biomarker for NSCLC.
Variants in IL 6ST selectively IL – 11 signal defects in the Human and mouse Gp130 cytokine receptor subunit encoded by the IL 6ST is a cytokine receptors together for ten of IL -6 family. We describe non-identical homozygous variant in IL 6ST (p.R281Q) in patients with craniosynostosis and maintain primary teeth. We characterize the effects of variants in cytokine signaling in vitro using cell lines and cells transfected patient-derived primary and support these findings using mice models with appropriate genome -edited variant Il6st that p.R279Q.
We showed that Human p.R281Q gp130 associated with selective loss IL – 11 signals without affecting the IL – 6 IL -27, OSM, LIF, CT1, CLC, and CNTF signal. In mice Il6st p.R279Q reduce the size of garbage and cause synostosis facial and dental abnormalities. Effects on IL – 11 signaling induced by gp130 variants showed incomplete penetrance but phenocopies aspect IL 11 RA deficiencies in the human and mice. Our data indicate that genetically in pleiotropic cytokine receptor variants may have a very selective flaw.
