Circ_0000527 promotes the progression of retinoblastoma by regulating miR-646/LRP6 axis
Circ_0000527 promotes the progression of retinoblastoma by regulating miR-646/LRP6 axis
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DOI：
10.1186/s12935-020-01396-4
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发表时间：
2020-07-10
影响因子：
5.8
通讯作者：
Ding, Xiaoyan
中科院分区：
文献类型：
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作者：
Zhang, Li;Wu, Jie;Ding, Xiaoyan
研究背景研究证实，环状RNA（circular RNAs，circRNA）在多种恶性肿瘤中表达异常，并在调节肿瘤细胞恶性表型中发挥重要作用。方法采用实时荧光定量PCR（Real-time PCR，RT-PCR）检测视网膜母细胞瘤（retinoblastoma，RB）组织和细胞中circ_0000527和miR-646的表达。Western blot检测RB细胞中LRP 6的表达。分析circ_0000527表达与RB患者临床病理参数的关系。采用CCK-8试剂盒、流式细胞仪和Transwell法检测细胞增殖、凋亡和转移情况。结果Circ_0000527在RB和RB细胞株中均高表达，且其高表达水平和分化程度与cTNM分期水平的增高显著相关。过表达circ_0000527有助于RB细胞的增殖、迁移、侵袭和抑制细胞凋亡，而敲低circ_0000527则抑制上述恶性生物学行为。结论Circ_0000527通过调节miR-646/LRP 6轴，促进RB细胞的增殖和转移。
BackgroundResearches 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.MethodsReal-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.ResultsCirc_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.ConclusionCirc_0000527 enhances the proliferation and metastasis of RB cells by modulating the miR-646/LRP6 axis.