NLK-shRNA Inhibits Tumor Cell Growth In H1299 Lung Adenocarcinoma Cell Line

Objective:Aberration of cell cycle regulation and cell proliferation are considered to be the main cause of malignant tumors. Previous studies have indicated that nemo-like kinase (NLK) is closely related to cell proliferation in different kinds of human cancers. However, the underlying mechanism is still unclear. The aim of current study is to confirm whether targeted disruption of NLK would inhibit cell proliferation and promote apoptosis of NSCLC cells, which would provide scientific basis for a further understanding of the development and progression of molecular mechanisms of NSCLC.Methods:Expression of NLK in five NSCLC cell lines (A549, H1299, NCI-H1581, NCI-H522 and SK-MES-1) was measured by westernblotting. Among them H1299 cell line was infected by NLK-shRNA lentiviral vector as experimental model. And real-time PCR was performed to detect the mRNA expression of NLK after infection. Cell proliferation assay, colony formation assay, flow cytometry cell cycle analysis and apoptosis experiments were used to analyze the alterations of NSCLC biological behaviors.Results: NLK are stable expressed in all five NSCLC cell lines (A549, H1299, NCI-1581, NCI-522 and SK-MES-1) measured by westernblotting, real-time PCR results showed that endogenous NLK was disrupted by NLK-shRNA lentiviral vector. Cell proliferation assay indicated that cell proliferation were suppressed, colony formation assay showed that NLK knockdown led to slow colony formation, flow cytometry cell cycle analysis results revealed significant Gl arrest in cell cycle and apoptosis experiments confirm more tumor cells were induced to apoptosis after infection.Conclusions:Aberrant expression of NLK might participate in the malignant progression of NSCLC.