Liu Zhenhua, Lv Chunye
Department of General Surgery, The First People's Hospital of Lin Ping District, Hangzhou, Zhejiang 311100, P.R. China.
Department of General Surgery, The Affiliated Jiangning Hospital with Nanjing Medical University, Nanjing, Jiangsu 211100, P.R. China.
Oncol Lett. 2022 Aug 17;24(4):346. doi: 10.3892/ol.2022.13466. eCollection 2022 Oct.
Pumilio homolog 2 (PUM2) is an RNA-binding protein that functions as an oncogene in various types of cancer. However, its role in hepatocellular carcinoma (HCC) has remained to be fully elucidated. In the present study, the role of PUM2 was investigated in HCC and its regulation was assessed by examining its binding to the 3'-untranslated region (UTR) of B-cell translocation gene 3 (BTG3). The expression levels of PUM2 were determined in datasets from the UALCAN and Cancer Cell Line Encyclopedia databases. Furthermore, Gene Expression Profiling Interactive Analysis was used to analyze overall survival in patients with HCC. Reverse transcription-quantitative PCR (RT-qPCR) and western blot analyses were then performed to detect the expression levels of PUM2 and BTG3 in HCC cells. Cell proliferation was assessed using Cell Counting Kit-8 and colony-formation assays. The induction of cell apoptosis was evaluated using TUNEL and western blotting assays. StarBase and RNA-Protein Interaction Prediction were used to determine the possible direct interaction between PUM2 and BTG3. The interaction between PUM2 and BTG3 was then verified by luciferase reporter and RNA-binding protein immunoprecipitation assays. The results indicated that PUM2 expression was upregulated in HCC tissues and cells and that it was associated with the prognosis of patients with HCC. PUM2 silencing inhibited the proliferation and promoted the apoptosis of Huh-7 cells. In addition, PUM2 was confirmed to directly bind to the 3'UTR of BTG3. Downregulation of BTG3 reversed the effects of PUM2 silencing on cell proliferation and apoptosis in Huh-7 cells. Collectively, the results suggested that PUM2 regulated HCC cell proliferation and apoptosis via interacting with BTG3, which may provide a novel therapeutic strategy for the treatment of human HCC.
Pumilio同源物2(PUM2)是一种RNA结合蛋白,在多种类型的癌症中发挥癌基因的作用。然而,其在肝细胞癌(HCC)中的作用仍有待充分阐明。在本研究中,研究了PUM2在HCC中的作用,并通过检测其与B细胞易位基因3(BTG3)的3'非翻译区(UTR)的结合来评估其调控机制。在来自UALCAN和癌细胞系百科全书数据库的数据集中测定了PUM2的表达水平。此外,使用基因表达谱交互式分析来分析HCC患者的总生存期。然后进行逆转录定量PCR(RT-qPCR)和蛋白质印迹分析,以检测HCC细胞中PUM2和BTG3的表达水平。使用细胞计数试剂盒-8和集落形成试验评估细胞增殖。使用TUNEL和蛋白质印迹试验评估细胞凋亡的诱导情况。使用StarBase和RNA-蛋白质相互作用预测来确定PUM2和BTG3之间可能的直接相互作用。然后通过荧光素酶报告基因和RNA结合蛋白免疫沉淀试验验证PUM2和BTG3之间的相互作用。结果表明,PUM2在HCC组织和细胞中表达上调,并且与HCC患者的预后相关。PUM2沉默抑制了Huh-7细胞的增殖并促进了其凋亡。此外,证实PUM2直接与BTG3的3'UTR结合。BTG3的下调逆转了PUM2沉默对Huh-7细胞增殖和凋亡的影响。总体而言,结果表明PUM2通过与BTG3相互作用来调节HCC细胞的增殖和凋亡,这可能为人类HCC的治疗提供一种新的治疗策略。