Exosomal circPACRGL promotes progression of colorectal cancer via the miR-142-3p/miR-506-3p-TGF-β1 axis
Exosomal circPACRGL promotes progression of colorectal cancer via the miR-142-3p/miR-506-3p-TGF-β1 axis
复制标题
外泌体 circPACRGL 通过 miR-142-3p/miR-506-3p-TGF-beta 1 轴促进结直肠癌的进展
DOI：
10.1186/s12943-020-01235-0
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发表时间：
2020-07-27
期刊：
影响因子：
37.3
通讯作者：
Li, Dong
中科院分区：
文献类型：
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作者：
Shang, Anquan;Gu, Chenzheng;Li, Dong
结直肠癌（CRC）是世界范围内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，并且大量的环状rna （circRNAs）在外泌体中富集。CircRNAs是调节癌症增殖和进展的非编码rna的新成员。然而，癌源性外泌体环状rna在结直肠癌中的功能和调控机制尚不清楚。方法采用透射电镜、纳米颗粒跟踪分析（NTA）和western blot对结直肠癌细胞来源的外泌体进行表征。CCK-8、伤口愈合和transwell检测以及流式细胞术检测分别评估外泌体是否会影响结直肠癌细胞的增殖、转移和凋亡。此外，我们进行了RNA测序和RT-qPCR来鉴定外泌体刺激的CRC细胞中的环状RNA。采用荧光原位杂交法（FISH）检测cirpacrgl的细胞分布。利用生物信息学分析（StarBase 2.0）对cirpacrgl的miRNA靶点进行汇总。荧光素酶测定证实直接相互作用。最后用流式细胞术检测N1-N2中性粒细胞的分化情况。我们的研究鉴定了一种新的源自crc的外泌体环状rna circPACRGL。我们发现，在肿瘤源性外泌体添加后，环pacrgl在结直肠癌细胞中显著上调。此外，circPACRGL作为miR-142-3p/miR-506-3p的海绵，促进转化生长因子- β 1（tgf - β 1）的表达。因此，circPACRGL通过miR-142-3p/ mir -506-3p- tgf - β 1轴促进结直肠癌细胞的增殖、迁移和侵袭，以及N1向N2中性粒细胞的分化。我们的研究首次揭示了癌源性外泌体cirpacrgl在CRC增殖和转移中起致癌作用，为circRNAs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记物。
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-beta 1(TGF-beta 1) 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-beta 1axis.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.