Effects of ANCR lncRNA on the biological behaviors of lung cancer cells A549 and the mechanism

TRANSLATIONAL CANCER RESEARCH(2020)

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摘要
Background: ANCR lncRNA has been reported to participate in many cancers, but its role in lung cancer remains unclear. FOXO1 is involved in the growth inhibition and prognosis of lung cancer. We aimed to evaluate the effects of ANCR lncRNA on the biological behaviors of lung cancer cells and the mechanism. Methods: The lung cancer and paracancerous tissues of 50 patients were collected. The expression levels of ANCR lncRNA and FOXO1 were detected by RT-qPCR and subjected to Pearson's correlation analysis. The correlations between ANCR Inc RNA expression and clinicopathological features of lung cancer patients were analyzed. The interaction between FOXO1 and ANCR lncRNA was studied by RNA pull-down and RNA immunoprecipitation assays. Human lung cancer A549 cells were transfected with lentiviral plasmids packaging ANCR lncRNA and FOXO1. The cells were divided into control group (untransfected), NC group (transfected with empty lentiviral plasmid), ANCR lncRNA group (transfected with pHRi-ANCR lncRNA), sh-ANCR lncRNA group (transfected with pHRi-sh-ANCR lncRNA), FOXO1 group (transfected with pHRi-FOXO1) and sh-ANCR lncRNA + sh-FOXO1 group (transfected with pHRi-sh-ANCR lncRNA and pI IRi-sh-FOXO1). The expression levels of FOXO1 after ANCR lncRNA overexpression or inhibition were measured by RT-qPCR. Cell proliferation, cell cycle and apoptosis were detected with 5-ethynyl-2'deoxyuridine labeling and flow cytometry respectively. Bcl-2, Bax, cyclin D1 and P27 protein expressions were detected by Western blot. Results: Compared with paracancerous tissue, ANCR expression in lung cancer tissue significantly increased, and FOXO1 significantly decreased (P<0.01), being negatively correlated (P<0.01). The patients with higher TNM stage, lower differentiation degree and lymph node metastasis had higher relative expression of ANCR lncRNA. ANCR lncRNA bound FOXO1 and inhibited its expression. Compared with the NC group, sh-ANCR lncRNA and FOXO1 groups had fewer proliferative cells, more significant arrest in the GO/G1 phase, more apoptotic cells, increased expressions of Bax and P27, and reduced expressions of Bcl-2 and cyclin D1 (P<0.05). Compared with the sh-ANCR lncRNA group, the above results of the shANCR lncRNA + sh-FOXO1 group were reversed (P<0.05). Conclusions: ANCR lncRNA had high expression in lung cancer, and regulated the proliferation and apoptosis of lung cancer cells by modulating FOXO1 expression.
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关键词
Long non-coding RNA (lncRNA),antidifferentiation noncoding RNA (ANCR),lung cancer,proliferation,cell cycle,apoptosis
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