J Zhang1, M Zhao, Z-Q Xue, Y Liu, Y-X Wang. 1. Department of Thoracic Surgery, Chinese PLA General Hospital, Haiding, Beijing, China. wydc1991@yeah.net.
Abstract
OBJECTIVE: This study is aimed to investigate the cellular role of miR-377 and demonstrate that miR-377 negatively regulated cyclin-dependent kinase 6(CDK6) in human non-small cell lung cancer (NSCLC) cells. MATERIALS AND METHODS: qRT-PCR was performed to identify the miR-377 expression level in 45 paired NSCLC and adjacent normal lung tissues. Cell proliferation was measured by MTT. Apoptosis was detected by flow cytometric analysis. Luciferase reporter assays were employed to validate regulation of a putative target of miR-377. The effect of miR-377 on endogenous levels of this target was subsequently confirmed via Western blot. RESULTS: We found that the expression level of miR-377 was significantly reduced in NSCLC tissues and cell lines. On the contrary, CDK6 expression level was up-regulated in NSCLC tissues and cell lines. Based on Luciferase reporter assays, we confirmed that CDK6 was a direct target gene of miR-377. In vitro studies demonstrated that miR-377 overexpression reduced NSCLC cell proliferation and promoted apoptosis. CONCLUSIONS: Our discovery suggested that miR-377 might be used as a therapeutic reagent for the treatment of NSCLC in the future.
OBJECTIVE: This study is aimed to investigate the cellular role of miR-377 and demonstrate that miR-377 negatively regulated cyclin-dependent kinase 6(CDK6) in humannon-small cell lung cancer (NSCLC) cells. MATERIALS AND METHODS: qRT-PCR was performed to identify the miR-377 expression level in 45 paired NSCLC and adjacent normal lung tissues. Cell proliferation was measured by MTT. Apoptosis was detected by flow cytometric analysis. Luciferase reporter assays were employed to validate regulation of a putative target of miR-377. The effect of miR-377 on endogenous levels of this target was subsequently confirmed via Western blot. RESULTS: We found that the expression level of miR-377 was significantly reduced in NSCLC tissues and cell lines. On the contrary, CDK6 expression level was up-regulated in NSCLC tissues and cell lines. Based on Luciferase reporter assays, we confirmed that CDK6 was a direct target gene of miR-377. In vitro studies demonstrated that miR-377 overexpression reduced NSCLC cell proliferation and promoted apoptosis. CONCLUSIONS: Our discovery suggested that miR-377 might be used as a therapeutic reagent for the treatment of NSCLC in the future.