为肝细胞癌武装 GPC3 CAR-T 细胞：多少才足够，下一步是什么？
Armouring GPC3 CAR T cells for hepatocellular carcinoma: how much is enough and what comes next?
CELL INTELLIGENCE · 肿瘤细胞治疗研究
肿瘤细胞治疗研究
Circ_0027791 通过 miR-496/程序性细胞死亡配体 1 轴以 m6A 依赖的方式促进肝细胞癌的生长和免疫逃逸
英文原题：Circ_0027791 contributes to the growth and immune evasion of hepatocellular carcinoma via the miR-496/programmed cell death ligand 1 axis in an m6A-dependent manner.
查看英文原题
Circ_0027791 contributes to the growth and immune evasion of hepatocellular carcinoma via the miR-496/programmed cell death ligand 1 axis in an m6A-dependent manner.
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中文摘要
新出现的证据表明，环状RNA在包括肝细胞癌（HCC）在内的多种癌症的发生发展中发挥关键作用。在此，我们目前的研究报道了circ_0027791在HCC进展中的生物学功能和机制。通过实时定量聚合酶链反应（RT-qPCR）检测circ_0027791、microRNA-496（miR-496）、程序性细胞死亡配体1（PDL1）和甲基转移酶样3（METTL3）的水平。使用3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四唑溴化物、5-乙炔基-2'-脱氧尿苷、transwell和成球实验检测细胞活力、增殖、侵袭和成球能力。使用流式细胞术检测巨噬细胞极化。
为了了解circ_0027791在免疫逃逸中的作用，将HCC细胞在体外与外周血单核细胞或细胞因子诱导的杀伤（CIK）细胞共培养。应用异种移植小鼠模型评估circ_0027791在体内的功能。在使用circinteractome和miRDB进行预测后，基于双荧光素酶报告基因实验验证了miR-496与circ_0027791或PDL1之间的结合。使用甲基化RNA免疫沉淀（MeRIP）-qPCR、RIP-qPCR和RNA pull-down实验确定METTL3与circ_0027791之间的相互作用。在HCC患者和细胞中，circ_0027791、PDL1和METTL3表达上调，而miR-496降低。
此外，敲低circ_0027791可能抑制增殖、侵袭、成球、M2巨噬细胞极化和抗肿瘤免疫反应。敲低circ_0027791在体内抑制HCC肿瘤生长。在机制上，circ_0027791作为miR-496的海绵来增加PDL1表达。
此外，METTL3介导了circ_0027791的m6A甲基化并稳定其表达。METTL3诱导的circ_0027791部分通过调控miR-496/PDL1轴促进HCC细胞进展，为HCC提供了新的预后和治疗标志物。
展开英文摘要原文
Emerging evidence indicates the critical roles of circular RNAs in the development of multiple cancers, containing hepatocellular carcinoma (HCC).
Herein, our present research reported the biological function and mechanism of circ_0027791 in HCC progression. Circ_0027791, microRNA-496 (miR-496), programmed cell death ligand 1 (PDL1), and methyltransferase-like 3 (METTL3) levels were detected by real-time quantitative polymerase chain reaction (RT-qPCR). Cell viability, proliferation, invasion, and sphere formation ability were detected using 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide, 5-ethynyl-2'-deoxyuridine, transwell, and sphere formation assays. Macrophage polarization was detected using flow cytometry assay.
To understand the role of circ_0027791 during the immune escape, HCC cells were cocultured with peripheral blood mononuclear cells or cytokine-induced killer (CIK) cells in vitro. A xenograft mouse model was applied to assess the function of circ_0027791 in vivo.
After prediction using circinteractome and miRDB, the binding between miR-496 and circ_0027791 or PDL1 was validated based on a dual-luciferase reporter assay. Interaction between METTL3 and circ_0027791 was determined using methylated RNA immunoprecipitation (MeRIP)-qPCR, RIP-qPCR, and RNA pull-down assays. Circ_0027791, PDL1, and METTL3 expression were upregulated, and miR-496 was decreased in HCC patients and cells.
Moreover, circ_0027791 knockdown might repress proliferation, invasion, sphere formation, M2 macrophage polarization, and antitumor immune response. Circ_0027791 knockdown repressed HCC tumor growth in vivo. In mechanism, circ_0027791 functioned as a sponge for miR-496 to increase PDL1 expression.
In addition, METTL3 mediated the m6A methylation of circ_0027791 and stabilized its expression. METTL3-induced circ_0027791 facilitated HCC cell progression partly regulating the miR-496/PDL1 axis, which provided a new prognostic and therapeutic marker for HCC.
论文信息
- 作者
- Yu F、Fang P、Fang Y、Chen D
- 单位
- Department of Medical Technology, Anhui Medical College, Hefei, China.China
- 期刊
- Environmental toxicology2024 Jun
- 原文标识
- PubMed 38546290 · DOI 10.1002/tox.24188