Antidifferentiation Noncoding RNA Regulates the Proliferation of Osteosarcoma Cells.

Abstract

Antidifferentiation noncoding RNA (ANCR), a newly identified long noncoding RNA (lncRNA), plays a critical role for stem cells to maintain undifferentiated cell state. However, the functions of ANCR in human cancers have not been reported. This study is designed to explore the role of ANCR in osteosarcoma. Lentivirus-mediated (shRNA) was applied to silence ANCR in the human osteosarcoma cell lines U2OS and Saos-2. Cell viability was measured by MTT assay. Colony-forming ability was measured by colony formation assay. Cell cycle progression was determined by flow cytometry with propidium iodide staining. In addition, cell cycle makers, including p21, CDK2, and CDK4, were investigated in ANCR silencing U2OS cells by real time PCR (RT-PCR) analysis. In this study, we first proved that lentivirus-mediated shRNA specifically suppressed the expression level of ANCR in U2OS and Saos-2 cells. Further investigations revealed that knockdown of ANCR significantly inhibited the proliferation of U2OS and Saos cells and colony formation of U2OS cells. Moreover, the cell cycle of U2OS cells was arrested at G0/G1 phase after ANCR knockdown. Furthermore, the expression level of p21 was increased and CDK2 was decreased in ANCR knock-down cells. Our data indicated that ANCR might be an oncogenic lncRNA that promoted proliferation of osteosarcoma. The potential application of ANCR-targeted therapy using the lentivirus-mediated shRNA approach is worth further investigations in preclinical and clinical studies.