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长链非编码RNA HOXA11-AS通过招募叉头框蛋白P2上调细胞周期蛋白D2,以抑制肾母细胞瘤细胞凋亡并促进细胞周期进程。

Long non-coding RNA HOXA11-AS upregulates Cyclin D2 to inhibit apoptosis and promote cell cycle progression in nephroblastoma by recruiting forkhead box P2.

作者信息

Zhu Shibo, Zhang Jingqi, Cui Yanhong, Tang Xiangliang, Gao Xiaofeng, Li Dian, Jia Wei

机构信息

The First Affiliated Hospital, Jinan University Guangzhou 510630, P. R. China.

Department of Pediatric Urology, Guangzhou Women and Children's Medical Center, Guangzhou Medical University Guangzhou 510623, P. R. China.

出版信息

Am J Cancer Res. 2020 Jan 1;10(1):284-298. eCollection 2020.

Abstract

Long non-coding RNAs (lncRNAs) have been highlighted to play key roles in the gene regulatory network, and the dysregulation of lncRNAs has also been implicated in various malignancies. However, little is known regarding the expression of lncRNA and their functions in the progression of nephroblastoma. Thus, the present study aimed to explore the potential role of homeobox A11 (HOXA11)-AS in nephroblastoma. Microarray-based analysis was initially applied to screen the differentially expressed lncRNAs, and HOXA11-AS was selected as the candidate. The HFWT cells were performed with gain- and loss-of function test to evaluate the role of HOXA11-AS in cell cycle and apoptosis in nephroblastoma using flow cytometry and Western blots. Moreover, the relationship between HOXA11-AS and forkhead box P2 (FOXP2) was verified by Cross-linking RIP, and the direct interaction between HOXA11-AS and Cyclin D2 (CCND2) was detected using a dual luciferase reporter gene assay. Tumor formation in nude mice was used to investigate the effect of HOXA11-AS . HOXA11-AS was found to be highly expressed in the nephroblastoma. Furthermore, the silencing of HOXA11-AS promoted apoptosis and cell cycle arrest at the G1/S phase in nephroblastoma through the transcription factor FOXP2 to downregulate the expression of CCND2. Consistently, the tumor formation data in nude mice verified the results . Taken together, silencing of HOXA11-AS promotes apoptosis and inhibits the cell cycle entry in nephroblastoma by recruiting the transcription factor FOXP2 to downregulate the expression of CCND2, highlighting a promising novel direction for future nephroblastoma treatment.

摘要

长链非编码RNA(lncRNAs)已被强调在基因调控网络中发挥关键作用,lncRNAs的失调也与多种恶性肿瘤有关。然而,关于lncRNA在肾母细胞瘤进展中的表达及其功能知之甚少。因此,本研究旨在探讨同源框A11(HOXA11)-AS在肾母细胞瘤中的潜在作用。最初应用基于微阵列的分析来筛选差异表达的lncRNAs,并选择HOXA11-AS作为候选者。对HFWT细胞进行功能获得和功能丧失测试,以使用流式细胞术和蛋白质免疫印迹评估HOXA11-AS在肾母细胞瘤细胞周期和凋亡中的作用。此外,通过交联RNA免疫沉淀(RIP)验证HOXA11-AS与叉头框P2(FOXP2)之间的关系,并使用双荧光素酶报告基因测定法检测HOXA11-AS与细胞周期蛋白D2(CCND2)之间的直接相互作用。利用裸鼠体内肿瘤形成来研究HOXA11-AS的作用。发现HOXA11-AS在肾母细胞瘤中高表达。此外,HOXA11-AS的沉默通过转录因子FOXP2促进肾母细胞瘤细胞凋亡并使细胞周期停滞在G1/S期,从而下调CCND2的表达。一致地,裸鼠体内肿瘤形成数据证实了该结果。综上所述,HOXA11-AS的沉默通过募集转录因子FOXP2下调CCND2的表达,促进肾母细胞瘤细胞凋亡并抑制细胞周期进入,为未来肾母细胞瘤治疗突出了一个有前景的新方向。

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