Knockdown of ZEB2-AS1 inhibits cell proliferation, invasion and induces apoptosis in osteosarcoma by combining with EZH2

B. Zhang, D.-B. Fan, L. Liu, Y. Qin, D.-Q. Feng

Department of Orthopedics, Shuyang Hospital of Traditional Chinese Medicine Affiliated to Nanjing University of Chinese Medicine, Suqian, China. zhangbinsy@126.com

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• Oncology

OBJECTIVE: This study aims to explore whether ZEB2-AS1 can promote the development of osteosarcoma by affecting the proliferation, invasion, and apoptosis of osteosarcoma cells.

PATIENTS AND METHODS: Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was performed to detect the ZEB2-AS1 expression in osteosarcoma tissue specimens and normal bone tissues. After ZEB2-AS1 downregulation, Cell Counting Kit-8 (CCK-8) test, plate cloning assay, 5-Ethynyl-2’-deoxyuridine (EdU) experiment, and flow cytometry were conducted to analyze the changes in cell proliferation and apoptosis. RIP assay was performed to detect the binding of ZEB2-AS1 to EZH2, while Western blot was applied to examine the EZH2 expression after EZH2 was inhibited. Meanwhile, after simultaneously inhibiting ZEB2-AS1 and EZH2, the cell invasiveness was determined by transwell assay.

RESULTS: ZEB2-AS1 was highly expressed in osteosarcoma tissues, especially in advanced and metastatic groups. Interfering with ZEB2-AS1 suppressed cell proliferation and enhanced cell apoptosis. In addition, ZEB2-AS1 was confirmed to be able to combine with EZH2. The knockdown of ZEB2-AS1 attenuated the cell invasion ability, which was further decreased after the simultaneous downregulation of ZEB2-AS1 and EZH2.

CONCLUSIONS: The long non-coding RNA, ZEB2-AS1, enhanced the proliferation and invasion of osteosarcoma cells and inhibited the cell apoptosis by combining with EZH2, and thereby promoted the development of osteosarcoma.