Zhang B, Fan D-B, Liu L, Qin Y, Feng D-Q
Department of Orthopedics, Shuyang Hospital of Traditional Chinese Medicine Affiliated to Nanjing University of Chinese Medicine, Suqian, China.
Eur Rev Med Pharmacol Sci. 2020 Jun;24(12):6533-6539. doi: 10.26355/eurrev_202006_21637.
This study aims to explore whether ZEB2-AS1 can promote the development of osteosarcoma by affecting the proliferation, invasion, and apoptosis of osteosarcoma cells.
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.
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.
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.
本研究旨在探讨ZEB2-AS1是否可通过影响骨肉瘤细胞的增殖、侵袭和凋亡来促进骨肉瘤的发展。
采用定量实时聚合酶链反应（qRT-PCR）检测骨肉瘤组织标本和正常骨组织中ZEB2-AS1的表达。在下调ZEB2-AS1后，进行细胞计数试剂盒-8（CCK-8）试验、平板克隆试验、5-乙炔基-2'-脱氧尿苷（EdU）实验和流式细胞术，以分析细胞增殖和凋亡的变化。进行RNA免疫沉淀（RIP）试验检测ZEB2-AS1与EZH2的结合情况，同时采用蛋白质免疫印迹法检测EZH2被抑制后其表达情况。此外，在同时抑制ZEB2-AS1和EZH2后，通过Transwell试验测定细胞侵袭能力。
ZEB2-AS1在骨肉瘤组织中高表达，尤其是在晚期和转移组。干扰ZEB2-AS1可抑制细胞增殖并增强细胞凋亡。此外，证实ZEB2-AS1能够与EZH2结合。下调ZEB2-AS1可减弱细胞侵袭能力，在同时下调ZEB2-AS1和EZH2后细胞侵袭能力进一步降低。
长链非编码RNA ZEB2-AS1通过与EZH2结合增强骨肉瘤细胞的增殖和侵袭能力，并抑制细胞凋亡，从而促进骨肉瘤的发展。
