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Reduced miR-363-3p expression in non-small cell lung cancer is associated with gemcitabine resistance via targeting of CUL4A.

OBJECTIVE: Accumulating evidence has suggested that aberrant expression of microRNAs (miRNAs) is associated with non-small cell lung cancer (NSCLC) proliferation, migration, invasion and chemotherapy resistance. Cullin4A (CUL4A) has been previously reported to desensitize NSCLC cells to chemotherapy treatment. However, whether miRNAs regulate CUL4A to promote chemotherapy resistance remains unknown.

PATIENTS AND METHODS: Tissues were obtained from 40 NSCLC patients who received surgery at the Yancheng City No. 1 People's Hospital. Cell Counting Kit-8 (CCK-8) assays were applied for the detection of cell proliferation; mRNA and protein levels were determined by Real Time-quantitative Polymerase Chain Reaction (RT-qPCR) and Western blot, respectively. The interaction between mRNA 3'UTR and miRNA was predicted by TargetScan and verified by Dual-Luciferase reporter assay.

RESULTS: In the present study, miR-363-3p levels were revealed to be significantly decreased in tumor tissues obtained from NSCLC patients compared with adjacent normal tissues. The results of the CCK-8 assays showed that the overexpression of miR-363-3p may slightly inhibit the proliferation of A549 and H23 cells. Notably, the transfection with miR-363-3p antagonists reduced the sensitivity of A549 and H23 cells to gemcitabine treatment, whereas the overexpression of miR-363-3p markedly increased the sensitivity of A549 and H23 cells to gemcitabine treatment. Furthermore, CUL4A mRNA and protein levels were revealed to be decreased in A549 cells transfected with miR-363-3p mimics. The Dual-Luciferase reporter assay results further suggested that CUL4A represents a target gene of miR-363-3p.

CONCLUSIONS: The results indicated that decreased miR-363-3p expression enhanced gemcitabine resistance in NSCLC cells via regulation of CUL4A.

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