Department of Pediatric Ophthalmology, The Second Affiliated Hospital of Nanchang University, Nanchang 330006, P.R. China.
Department of Pediatric Ophthalmology, The Second Affiliated Hospital of Nanchang University, Nanchang 330006, P.R. China
Biosci Rep. 2018 Dec 11;38(6). doi: 10.1042/BSR20180558. Print 2018 Dec 21.
As one of the most common primary intraocular carcinomas, retinoblastoma generally stems from the inactivation of the retinoblastoma gene in retinal cells. Antisense non-coding RNA in the INK4 locus (ANRIL), a long non-coding RNA (lncRNA), has been reported to affect tumorigenesis and progression of various cancers, including gastric cancer and non-small cell lung cancer. However, limited investigations emphasized the role of ANRIL in human retinoblastoma. Hence, the current study was intended to investigate the effects of ANRIL on the proliferation, apoptosis, and invasion of retinoblastoma HXO-RB and Y79 cells. The lentivirus-based packaging system was designed to aid the up-regulation of ANRIL and ATM expressions or employed for the down-regulation of ANRIL in human retinoblastoma cells. Afterward, ANRIL expression, mRNA and protein expression of ATM and E2F1, and protein expression of INK4b, INK4a, alternate reading frame (ARF), p53 and retinoblastoma protein (pRB) were determined in order to elucidate the regulation effect associated with ANRIL on the ATM-E2F1 signaling pathway. In addition, cell viability, apoptosis, and invasion were detected accordingly. The results indicated that the down-regulation of ANRIL or up-regulation of ATM led to an increase in the expressions of ATM, E2F1, INK4b, INK4a, ARF, p53, and pRB. The silencing of ANRIL or up-regulation of ATM exerted an inhibitory effect on the proliferation and invasion while improving the apoptosis of HXO-RB and Y79 cells. In conclusion, the key observations of our study demonstrated that ANRIL depletion could act to suppress retinoblastoma progression by activating the ATM-E2F1 signaling pathway. These results provide a potentially promising basis for the targetted intervention treatment of human retinoblastoma.
作为最常见的原发性眼内癌之一，视网膜母细胞瘤通常源于视网膜细胞中视网膜母细胞瘤基因的失活。INK4 基因座中的反义非编码 RNA（ANRIL）是一种长链非编码 RNA（lncRNA），已被报道影响包括胃癌和非小细胞肺癌在内的多种癌症的发生和进展。然而，有限的研究强调了 ANRIL 在人视网膜母细胞瘤中的作用。因此，本研究旨在研究 ANRIL 对人视网膜母细胞瘤 HXO-RB 和 Y79 细胞增殖、凋亡和侵袭的影响。基于慢病毒的包装系统被设计用于上调 ANRIL 和 ATM 的表达，或用于下调人视网膜母细胞瘤细胞中的 ANRIL。随后，确定 ANRIL 表达、ATM 和 E2F1 的 mRNA 和蛋白表达以及 INK4b、INK4a、交替阅读框（ARF）、p53 和视网膜母细胞瘤蛋白（pRB）的蛋白表达，以阐明与 ANRIL 相关的对 ATM-E2F1 信号通路的调节作用。此外，相应地检测细胞活力、凋亡和侵袭。结果表明，下调 ANRIL 或上调 ATM 导致 ATM、E2F1、INK4b、INK4a、ARF、p53 和 pRB 的表达增加。沉默 ANRIL 或上调 ATM 对 HXO-RB 和 Y79 细胞的增殖和侵袭产生抑制作用，同时改善细胞凋亡。总之，我们研究的主要观察结果表明，通过激活 ATM-E2F1 信号通路，ANRIL 的缺失可以抑制视网膜母细胞瘤的进展。这些结果为人类视网膜母细胞瘤的靶向干预治疗提供了潜在的有希望的基础。
