Department of Hematology, The First Affiliated Hospital of Jinzhou Medical University, Jinzhou, Liaoning 121001, P.R. China.
Department of Geratology, The First Affiliated Hospital of Jinzhou Medical University, Jinzhou, Liaoning 121001, P.R. China.
Mol Med Rep. 2021 Feb;23(2). doi: 10.3892/mmr.2020.11785. Epub 2020 Dec 16.
Burkitt lymphoma (BL) has a high mortality rate and its treatment is currently limited to chemotherapy combined with immunotherapy. The long non‑coding RNA antisense non‑coding RNA in the INK4 locus (ANRIL) has been identified as an oncogene that can regulate cell proliferation and apoptosis in multiple types of cancer. However, the function of ANRIL in BL remains unknown. The present study aimed to determine the effect of ANRIL on cell proliferation and apoptosis in BL. Reverse transcription‑quantitative PCR was used to analyze the expression levels of ANRIL in BL cells. The effect of ANRIL knockdown on BL cells was determined using Cell Counting Kit‑8, flow cytometric, western blotting, immunofluorescence staining and Hoechst staining assays. The results revealed that ANRIL silencing inhibited the proliferation and promoted the apoptosis of BL cells. In addition, the expression levels of cyclin D1, E2F transcription factor 1 and Bcl‑2 were downregulated, while the expression levels of cyclin‑dependent kinase inhibitor 1A, Bcl‑2‑associated X protein, cleaved‑caspase‑9/pro‑caspase‑9 and cleaved‑caspase‑3/pro‑caspase‑3 were upregulated. Furthermore, the knockdown of ANRIL activated the TGF‑β1 signaling pathway, as evidenced by the upregulated expression levels of TGF‑β1, phosphorylated (p)‑SMAD2/3/SMAD2/3, p‑SMAD1/SMAD1 and sphingosine‑1‑phosphate receptor 2. Moreover, the protective effect of ANRIL silencing in BL could be inhibited by the TGF‑β receptor type I/II dual inhibitor, LY2109761. In conclusion, the findings of the present study suggested that the knockdown of ANRIL may inhibit cell proliferation and promote cell apoptosis in BL by regulating the TGF‑β1 signaling pathway, which may provide a novel target for the treatment of BL.
伯基特淋巴瘤(BL)死亡率高,目前的治疗方法仅限于化疗联合免疫疗法。已鉴定出位于 INK4 基因座的长链非编码 RNA 反义非编码 RNA(ANRIL)是一种癌基因,可调节多种类型癌症中的细胞增殖和凋亡。然而,ANRIL 在 BL 中的功能尚不清楚。本研究旨在确定 ANRIL 对 BL 细胞增殖和凋亡的影响。采用逆转录-定量 PCR 分析 BL 细胞中 ANRIL 的表达水平。通过细胞计数试剂盒-8、流式细胞术、Western blot 印迹、免疫荧光染色和 Hoechst 染色检测 ANRIL 敲低对 BL 细胞的影响。结果显示,ANRIL 沉默抑制 BL 细胞增殖并促进其凋亡。此外,下调细胞周期蛋白 D1、E2F 转录因子 1 和 Bcl-2 的表达水平,而上调细胞周期蛋白依赖性激酶抑制剂 1A、Bcl-2 相关 X 蛋白、裂解-胱天蛋白酶-9/原胱天蛋白酶-9 和裂解-胱天蛋白酶-3/原胱天蛋白酶-3 的表达水平。此外,ANRIL 敲低激活 TGF-β1 信号通路,表现为 TGF-β1、磷酸化 SMAD2/3/SMAD2/3、p-SMAD1/SMAD1 和鞘氨醇 1-磷酸受体 2 的表达水平上调。此外,TGF-β 受体 I/II 双重抑制剂 LY2109761 可抑制 ANRIL 沉默对 BL 的保护作用。综上所述,本研究结果表明,通过调节 TGF-β1 信号通路,ANRIL 敲低可能抑制 BL 细胞增殖并促进细胞凋亡,这可能为 BL 的治疗提供新的靶点。