环状 RNA hsa_circ_0000282 通过调控 miR-192/XIAP 轴促进骨肉瘤细胞增殖。
Circular RNA hsa_circ_0000282 contributes to osteosarcoma cell proliferation by regulating miR-192/XIAP axis.
机构信息
Department of Spine Surgery, Honghui Hospital, Xi'an Jiaotong University, No. 76 Nanguo Road, Xi'an, 710054, Shaanxi, China.
出版信息
BMC Cancer. 2020 Oct 23;20(1):1026. doi: 10.1186/s12885-020-07515-8.
BACKGROUND
Circular RNAs (circRNAs) have emerged as a novel category of non-coding RNA, which exhibit a pivotal effect on regulating gene expression and biological functions, yet how circRNAs function in osteosarcoma (OSA) still demands further investigation. This study aimed at probing into the function of hsa_circ_0000282 in OSA.
METHODS
The expressions of circ_0000282 and miR-192 in OSA tissues and cell lines were examined by quantitative real-time polymerase chain reaction (qRT-PCR), and the correlation between the expression level of circ_0000282 and clinicopathological features of OSA patients was analyzed. The expressions of X-linked inhibitor of apoptosis protein (XIAP), B-cell lymphoma-2 (Bcl-2) and Bcl-2 associated X protein (Bax) in OSA cells were assayed by Western blot. The proliferation and apoptosis of OSA cells were examined by CCK-8, BrdU and flow cytometry, respectively. Bioinformatics analysis, dual-luciferase reporter gene assay and RIP experiments were employed to predict and validate the targeting relationships between circ_0000282 and miR-192, and between miR-192 and XIAP, respectively.
RESULTS
Circ_0000282 was highly expressed in OSA tissues and cell lines, which represented positive correlation with Enneking stage of OSA patients and negative correlation with tumor differentiation degree. In vitro experiments confirmed that overexpression of circ_0000282 markedly facilitated OSA cell proliferation and repressed cancer cell apoptosis in comparison to control group. Besides, knockdown of circ_0000282 repressed OSA cell proliferation and promoted apoptosis. Additionally, the binding relationships between circ_0000282 and miR-192, and between miR-192 and XIAP were validated. Circ_0000282 indirectly up-regulated XIAP expression by adsorbing miR-192, thereby playing a role in promoting cancer in OSA.
CONCLUSION
Circ_0000282 was a novel oncogenic circRNA in OSA. Circ_0000282/miR-192/XIAP axis regulated OSA cell proliferation apoptosis with competitive endogenous RNA mechanism.
背景
环状 RNA（circRNAs）作为一种新型非编码 RNA，在调控基因表达和生物学功能方面发挥着关键作用，但 circRNAs 在骨肉瘤（OSA）中的作用仍有待进一步研究。本研究旨在探讨 hsa_circ_0000282 在 OSA 中的功能。
方法
采用实时定量聚合酶链反应（qRT-PCR）检测 OSA 组织和细胞系中 circ_0000282 和 miR-192 的表达，并分析 circ_0000282 表达水平与 OSA 患者临床病理特征的相关性。Western blot 检测 OSA 细胞中 X 连锁凋亡抑制蛋白（XIAP）、B 细胞淋巴瘤-2（Bcl-2）和 Bcl-2 相关 X 蛋白（Bax）的表达。分别采用 CCK-8、BrdU 和流式细胞术检测 OSA 细胞的增殖和凋亡。采用生物信息学分析、双荧光素酶报告基因检测和 RIP 实验分别预测和验证 circ_0000282 与 miR-192、miR-192 与 XIAP 之间的靶向关系。
结果
circ_0000282 在 OSA 组织和细胞系中高表达，与 OSA 患者的 Enneking 分期呈正相关，与肿瘤分化程度呈负相关。体外实验证实，与对照组相比，circ_0000282 的过表达显著促进 OSA 细胞增殖，抑制癌细胞凋亡。此外，circ_0000282 的敲低抑制了 OSA 细胞的增殖并促进了凋亡。此外，还验证了 circ_0000282 与 miR-192 之间以及 miR-192 与 XIAP 之间的结合关系。circ_0000282 通过吸附 miR-192 间接上调 XIAP 表达，从而在 OSA 中发挥促进癌症的作用。
结论
circ_0000282 是 OSA 中的一种新型致癌 circRNA。circ_0000282/miR-192/XIAP 轴通过竞争性内源性 RNA 机制调节 OSA 细胞增殖凋亡。
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