Department of Pathology, The Second Affiliated Hospital of Harbin Medical University, Heilongjiang. China.
Department of Stomatology, The Second Affiliated Hospital of Harbin Medical University. Harbin Institute of Technology, Heilongjiang Provincial Hospital, Heilongjiang, China.
Bioengineered. 2021 Dec;12(1):5085-5098. doi: 10.1080/21655979.2021.1955561.
Long non-coding RNAs (lncRNAs) have been proposed as potential targets in OSCC gene therapy. Thus, the study aims to analyze how they exert functions in OSCC. LINC00958, AIM2, Gasdermin D (GSDMD) and tumor protein p53 (TP53) expression levels are analyzed by Quantitative Real-time PCR (qPCR) or Western blotting (WB) in OSCC cells lines. The roles of LINC00958 in cell proliferation, cell death, and GSDMD expression respectively were analyzed by Cell Counting Kit-8 (CCK8) assay, flow cytometry and Immunofluorescence (IF) assay. In addition, expressions of pyroptosis- and autophagy-related proteins are evaluated by WB detection. The targeted binding of LINC00958 and miR-4306 or AIM2 mRNA is predicted by bioinformatics analysis and detected by biodual luciferase system. RIP and qPCR assays analyze whether LINC00958 interacts with SIRT1. We found that LINC00958 showed upregulation in OSCC cells compared to normal oral epithelial cells. LINC00958 silencing significantly suppressed OSCC cell proliferation, induced cell death and reduced autophagy. LINC00958 regulated the levels of miR-4306 which binds to the 3'UTR of AIM2, and interacts with and modulates SIRT1 protein expression. LINC00958 regulated GSDMD and AIM2 levels, as well as p53 and SIRT1 levels. SIRT1 overexpression markedly reversed aforementioned effects of LINC00958. LINC00958 not only downregulated miR-4306 levels to activate the pyroptosis pathway mediated by AIM2 and promoted cancer cell survival but also induced a decrease in SIRT protein expression to further reduce p53 levels.
长链非编码 RNA(lncRNA)已被提出作为口腔鳞状细胞癌(OSCC)基因治疗的潜在靶点。因此,本研究旨在分析它们在 OSCC 中发挥功能的方式。通过定量实时 PCR(qPCR)或 Western blot(WB)分析 OSCC 细胞系中的 LINC00958、AIM2、Gasdermin D(GSDMD)和肿瘤蛋白 p53(TP53)的表达水平。通过细胞计数试剂盒-8(CCK8)测定、流式细胞术和免疫荧光(IF)测定分别分析 LINC00958 对细胞增殖、细胞死亡和 GSDMD 表达的作用。此外,通过 WB 检测评估细胞焦亡和自噬相关蛋白的表达。通过生物信息学分析预测 LINC00958 与 miR-4306 或 AIM2 mRNA 的靶向结合,并通过双荧光素酶系统检测。RIP 和 qPCR 测定分析 LINC00958 是否与 SIRT1 相互作用。我们发现,与正常口腔上皮细胞相比,LINC00958 在 OSCC 细胞中上调。LINC00958 沉默显著抑制 OSCC 细胞增殖,诱导细胞死亡并减少自噬。LINC00958 调节 miR-4306 的水平,miR-4306 结合 AIM2 的 3'UTR,并与 SIRT1 蛋白相互作用并调节其表达。LINC00958 调节 GSDMD 和 AIM2 水平以及 p53 和 SIRT1 水平。SIRT1 的过表达显著逆转了 LINC00958 的上述作用。LINC00958 不仅通过下调 miR-4306 水平来激活由 AIM2 介导的细胞焦亡途径,促进癌细胞存活,还通过诱导 SIRT 蛋白表达减少来进一步降低 p53 水平。