环状RNA_0000527通过调控miR-646/LRP6轴促进视网膜母细胞瘤进展。
Circ_0000527 promotes the progression of retinoblastoma by regulating miR-646/LRP6 axis.
作者信息
Zhang Li, Wu Jie, Li Yujun, Jiang Yanxia, Wang Lili, Chen Yunqing, Lv Yalin, Zou Yuwei, Ding Xiaoyan
机构信息
Department of Pathology, The Affiliated Hospital of Qingdao University, Qingdao, 266003 Shandong China.
Department of Dermatology, The Affiliated Hospital of Qingdao University, No 16, Jiangsu Road, South District, Qingdao, 266003 Shandong China.
出版信息
Cancer Cell Int. 2020 Jul 10;20:301. doi: 10.1186/s12935-020-01396-4. eCollection 2020.
BACKGROUND
Researches validate that circular RNAs (circRNAs) are dysregulated in a variety of malignancies and play an important role in regulating the malignant phenotype of tumor cells. Nevertheless, the role of circ_0000527 in retinoblastoma (RB) and its regulatory mechanisms remain largely unknown.
METHODS
Real-time PCR (RT-PCR) was used to detect circ_0000527 and miR-646 expression in RB tissues and cells. The LRP6 expression in RB cells was detected by Western blot. The relationship between circ_0000527 expression and the clinicopathological parameters of RB patients was analyzed. Cell proliferation, apoptosis and metastasis were monitored by cell counting kit-8 (CCK-8), flow cytometry, and Transwell assay. The dual-luciferase reporter gene assay and RIP assay were employed to verify the targeting relationship between circ_0000527 and miR-646, miR-646 and LRP6.
RESULTS
Circ_0000527 was highly expressed in both RB and RB cell lines, whose high expression level and degree of differentiation were significantly correlated with the increase in cTNM staging level. Overexpression of circ_0000527 contributed to RB cell proliferation, migration, invasion and suppressed cell apoptosis, while knocking down circ_0000527 inhibited the above malignant biological behavior. The underlying mechanism suggested that functioning as a endogenous competitive RNA, circ_0000527 directly targeted miR-646 and positively regulated LRP6 expression.
CONCLUSION
Circ_0000527 enhances the proliferation and metastasis of RB cells by modulating the miR-646/LRP6 axis.
背景
研究证实，环状RNA（circRNA）在多种恶性肿瘤中表达失调，并在调节肿瘤细胞的恶性表型中发挥重要作用。然而，circ_0000527在视网膜母细胞瘤（RB）中的作用及其调控机制仍不清楚。
方法
采用实时荧光定量PCR（RT-PCR）检测RB组织和细胞中circ_0000527和miR-646的表达。采用蛋白质免疫印迹法检测RB细胞中低密度脂蛋白受体相关蛋白6（LRP6）的表达。分析circ_0000527表达与RB患者临床病理参数之间的关系。采用细胞计数试剂盒-8（CCK-8）、流式细胞术和Transwell实验监测细胞增殖、凋亡和迁移。采用双荧光素酶报告基因实验和RNA免疫沉淀实验（RIP）验证circ_0000527与miR-646、miR-646与LRP6之间的靶向关系。
结果
circ_0000527在RB组织和细胞系中均高表达，其高表达水平及分化程度与cTNM分期水平升高显著相关。circ_0000527的过表达促进了RB细胞的增殖、迁移、侵袭并抑制细胞凋亡，而敲低circ_0000527则抑制了上述恶性生物学行为。潜在机制表明，circ_0000527作为内源性竞争RNA，直接靶向miR-646并正向调节LRP6的表达。
结论
circ_0000527通过调控miR-646/LRP6轴增强RB细胞的增殖和转移。
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