Jiangsu Institute of Clinical Immunology, The First Affiliated Hospital of Soochow University, Suzhou, China.
Department of Gastroenterology, The First Affiliated Hospital of Soochow University, Suzhou, China.
Cancer Sci. 2021 Jun;112(6):2245-2259. doi: 10.1111/cas.14896. Epub 2021 Apr 7.
Long noncoding RNAs (lncRNAs) have been investigated in multiple human cancers including gastric cancer (GC). Our research aims to explore the role of H19 in aerobic glycolysis, proliferation, and immune escape of GC cells. The expression of H19 in GC samples was analyzed using Gene Expression Profiling Interactive Analysis, Gene Expression Omnibus data, and real-time quantitative PCR analysis. Relative quantification of glucose consumption and lactate production from cell supernatant were applied to assess the aerobic glycolysis of GC cells. Subcellular fractionation, luciferase reporter, and western blot assays certified the binding between genes. Cell Counting Kit-8 and colony formation assays were used to determine GC cell proliferation. Flow cytometry, ELISA, and real-time quantitative PCR assays were applied to analyze the immunosuppressive effect of H19. H19 was highly expressed in samples of patients with GC, and associated with tumor growth in vivo. H19 knockdown suppressed glucose consumption, lactate production, and proliferation of GC cells by regulating the microRNA (miR)-519d-3p/lactate dehydrogenase A (LDHA) axis. Both miR-519d-3p depletion and LDHA overexpression could reverse the H19 knockdown-induced decrease in aerobic glycolysis and proliferation. Moreover, conditioned medium from stable knockdown H19 GC cells modulated the activity of immune cells including γδT cells, Jurkat cells, and tumor-associated macrophages in a miR-519d-3p/LDHA/lactate axis-dependent manner. The H19/miR-519d-3p/LDHA axis mainly contributed to aerobic glycolysis, proliferation, and immune escape of GC cells.
长链非编码 RNA（lncRNA）已在多种人类癌症中进行了研究，包括胃癌（GC）。我们的研究旨在探索 H19 在 GC 细胞的有氧糖酵解、增殖和免疫逃逸中的作用。使用基因表达谱分析交互分析、基因表达综合数据库数据和实时定量 PCR 分析来分析 GC 样本中 H19 的表达。应用细胞上清液中葡萄糖消耗和乳酸产生的相对定量来评估 GC 细胞的有氧糖酵解。亚细胞分离、荧光素酶报告和 Western blot 测定证明了基因之间的结合。细胞计数试剂盒-8 和集落形成测定用于确定 GC 细胞的增殖。流式细胞术、ELISA 和实时定量 PCR 测定用于分析 H19 的免疫抑制作用。H19 在 GC 患者的样本中高表达，并与体内肿瘤生长相关。H19 敲低通过调节 microRNA（miR）-519d-3p/乳酸脱氢酶 A（LDHA）轴来抑制 GC 细胞的葡萄糖消耗、乳酸产生和增殖。miR-519d-3p 耗竭和 LDHA 过表达均可逆转 H19 敲低诱导的有氧糖酵解和增殖减少。此外，稳定敲低 H19 的 GC 细胞条件培养基通过 miR-519d-3p/LDHA/乳酸轴依赖的方式调节免疫细胞的活性，包括 γδT 细胞、Jurkat 细胞和肿瘤相关巨噬细胞。H19/miR-519d-3p/LDHA 轴主要促进 GC 细胞的有氧糖酵解、增殖和免疫逃逸。
