外泌体 circPACRGL 通过 miR-142-3p/miR-506-3p-TGF-β1 轴促进结直肠癌的进展。
Exosomal circPACRGL promotes progression of colorectal cancer via the miR-142-3p/miR-506-3p- TGF-β1 axis.
机构信息
Department of Laboratory Medicine, Shanghai Tongji Hospital, Tongji University School of Medicine, No. 389, Xincun Road, Putuo District, Shanghai, 200065, P.R. China.
Department of Pathology, The Sixth People's Hospital of Yancheng City, Yancheng, 224001, P.R. China.
出版信息
Mol Cancer. 2020 Jul 27;19(1):117. doi: 10.1186/s12943-020-01235-0.
BACKGROUND
Colorectal cancer (CRC) is the leading cause of cancer-related death worldwide. Exosome shave emerged as crucial regulators of intercellular communication and that abundant Circular RNAs (circRNAs) are enriched within exosomes. CircRNAs are novel members of noncoding RNAs regulating cancer proliferation and progression. However, the function and regulatory mechanism of cancer-derived exosomal circRNAs in CRC remains unclear.
METHODS
CRC cells-derived exosomes were characterized using transmission electron microscopy, nanoparticle tracking analysis (NTA) and western blot. CCK-8, wound healing and transwell assays, and flow cytometry assays were conducted to assess whether exosomes would affect the proliferation, metastasis, and apoptosis of CRC cells, respectively. Moreover, we performed the RNA sequencing and RT-qPCR to identify circRNAs in exosome-stimulated CRC cells. Fluorescence in situ hybridization (FISH) assay was used to detect the cellular distribution of circPACRGL. Bioinformatic analyses (StarBase 2.0) were used to pool the miRNA targets of circPACRGL. Luciferase assays were performed to verify the direct interaction. Finally, flow cytometry was used to detect the differentiation of N1-N2 neutrophils.
RESULTS
Our study identified a novel CRC-derived exosomal circRNA, circPACRGL. We found circPACRGL was significantly upregulated in CRC cells after tumor-derived exosomes addition. Moreover, circPACRGL serves as a sponge for miR-142-3p/miR-506-3p to facilitate the transforming growth factor-β1 (TGF-β1) expression. As a result, circPACRGL promoted CRC cell proliferation, migration and invasion, as well as differentiation of N1 to N2 neutrophils via miR-142-3p/miR-506-3p-TGF-β1 axis.
CONCLUSION
Our study, the first to reveal that cancer-derived exosomal circPACRGL plays an oncogenic role in CRC proliferation and metastasis, providing mechanistic insights into the roles of circRNAs in CRC progression and a valuable marker for CRC treatment.
背景
结直肠癌（CRC）是全球癌症相关死亡的主要原因。外泌体已成为细胞间通讯的重要调节因子，大量环状 RNA（circRNA）富含于外泌体中。circRNA 是一种新型非编码 RNA，可调节癌症的增殖和进展。然而，CRC 中源自癌症的外泌体 circRNA 的功能和调节机制尚不清楚。
方法
使用透射电子显微镜、纳米颗粒跟踪分析（NTA）和 Western blot 对 CRC 细胞来源的外泌体进行表征。通过 CCK-8、划痕愈合和 Transwell 检测分别评估外泌体是否影响 CRC 细胞的增殖、转移和凋亡。此外，我们进行了 RNA 测序和 RT-qPCR 以鉴定外泌体刺激的 CRC 细胞中的 circRNA。荧光原位杂交（FISH）检测用于检测 circPACRGL 的细胞内分布。使用 StarBase 2.0 进行生物信息学分析以汇集 circPACRGL 的 miRNA 靶标。进行荧光素酶测定以验证直接相互作用。最后，使用流式细胞术检测 N1-N2 中性粒细胞的分化。
结果
我们鉴定出一种新型 CRC 来源的外泌体 circRNA，circPACRGL。我们发现，肿瘤衍生的外泌体添加后 CRC 细胞中的 circPACRGL 显著上调。此外，circPACRGL 作为 miR-142-3p/miR-506-3p 的海绵，促进转化生长因子-β1（TGF-β1）的表达。结果，circPACRGL 通过 miR-142-3p/miR-506-3p-TGF-β1 轴促进 CRC 细胞增殖、迁移和侵袭，并促进 N1 向 N2 中性粒细胞分化。
结论
我们的研究首次揭示了源自癌症的外泌体 circPACRGL 在 CRC 增殖和转移中发挥致癌作用，为 circRNA 在 CRC 进展中的作用提供了机制见解，并为 CRC 治疗提供了有价值的标志物。
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