Department of Oncology, Tongji Hospital, Tongji University School of Medicine, No. 389, Putuoxincun Rd., Shanghai, 200065, China.
Department of Colorectal Surgery, Department of General Surgery, Shanghai East Hospital, Tongji University School of Medicine, 150 Jimo Road, Shanghai, 200120, China.
J Nanobiotechnology. 2021 May 10;19(1):133. doi: 10.1186/s12951-021-00873-w.
Forkhead box protein M1 (FOXM1) is an oncogene regulating tumor growth and metastasis. Exosome was suggested to mediate cell communication by delivering active molecules in cancers. However, the existence of FOXM1 in circulating exosomes and the role of exosome FOXM1 in gastric cancer (GC) were not clear. This study aims to investigate the potential role of FOXM1 related long noncoding RNA (FRLnc1) in exosomes in GC.
The prepared CD63 immunomagnetic beads (CD63-IMB) had the characteristics of good dispersity and high magnetic response. The isolated exosomes were presented with elliptical membranous particles under a transmission electron microscope (TEM), with the particle size of 89.78 ± 4.8 nm. Western blot (WB) results showed that the exosomes were rich in CD9 and CD81. The Dil-labeled exosomes were distributed around cytoplasm and nucleus of cells by imaging flow cytometry (IFC) analysis. The results of quantitative real-time PCR (qRT-PCR) revealed that the FRLnc1 expressions were up-regulated in GC cells, tumor tissues, and serum of GC patients. An obviously up-regulated FRLnc1 expression was found in serum exosomes of GC patients. Up-regulation of FRLnc1 expression was closely correlated to lymph node metastasis (LNM) and TNM stage with the combination of relevant clinicopathological parameter analysis. The in vitro functional analyses demonstrated that FRLnc1 knockdown by RNA interference suppressed cell proliferation and migration in HGC-27 cells, whereas FRLnc1 overexpression promoted cell proliferation and migration in MKN45 cells. After exosome treatment, the FRLnc1 expression was significantly increased in MKN45 cells, and the MKN45 cells showed increased ability of proliferation and migration.
GC cells-derived exosomes played roles in promoting the growth and metastasis of GC by transporting FRLnc1, suggesting that FRLnc1 in the exosomes may be a potential biomarker for the diagnosis and treatment of GC. The delivery of FRLnc1 by the exosomes may provide a new way for the treatment of GC. Trial registration 2020-KYSB-094. Registered 23 March 2020-Retrospectively registered.
叉头框蛋白 M1（FOXM1）是一种调节肿瘤生长和转移的癌基因。外泌体被认为通过在癌症中传递活性分子来介导细胞间通讯。然而，FOXM1 在循环外泌体中的存在以及外泌体 FOXM1 在胃癌（GC）中的作用尚不清楚。本研究旨在探讨与 FOXM1 相关的长非编码 RNA（FRLnc1）在 GC 中外泌体中的潜在作用。
制备的 CD63 免疫磁珠（CD63-IMB）具有良好的分散性和高磁响应性。透射电子显微镜（TEM）下分离的外泌体呈椭圆形膜性颗粒，粒径为 89.78±4.8nm。Western blot（WB）结果显示外泌体富含 CD9 和 CD81。通过成像流式细胞术（IFC）分析发现，Dil 标记的外泌体分布在细胞的细胞质和核周围。实时荧光定量 PCR（qRT-PCR）结果显示，GC 细胞、肿瘤组织和 GC 患者血清中 FRLnc1 的表达上调。GC 患者血清外泌体中 FRLnc1 的表达明显上调。FRLnc1 表达上调与淋巴结转移（LNM）和 TNM 分期密切相关，结合相关临床病理参数分析。体外功能分析表明，RNA 干扰下调 FRLnc1 表达可抑制 HGC-27 细胞的增殖和迁移，而过表达 FRLnc1 可促进 MKN45 细胞的增殖和迁移。外泌体处理后，MKN45 细胞中 FRLnc1 的表达显著增加，MKN45 细胞增殖和迁移能力增强。
GC 细胞来源的外泌体通过转运 FRLnc1 促进 GC 的生长和转移，提示外泌体中的 FRLnc1 可能是 GC 诊断和治疗的潜在标志物。外泌体递送 FRLnc1 可能为 GC 的治疗提供新途径。试验注册 2020-KYSB-094。2020 年 3 月 23 日注册-回顾性注册。
