Exosomal circ_0084043 derived from colorectal cancer-associated fibroblasts promotes in vitro endothelial cell angiogenesis by regulating the miR-140-3p/HIF-1 α /VEGF signaling axis

被引:4
作者
Payervand, Nafiseh [1 ]
Pakravan, Katayoon [2 ]
Razmara, Ehsan [3 ,4 ]
Vinu, Kailash Kumar [4 ]
Ghodsi, Sara [2 ]
Heshmati, Masoumeh [1 ]
Babashah, Sadegh [2 ]
机构
[1] Islamic Azad Univ, Fac Adv Sci & Technol, Dept Cellular & Mol Biol, Tehran Med Sci, Tehran, Iran
[2] Tarbiat Modares Univ, Fac Biol Sci, Dept Mol Genet, Tehran, Iran
[3] Tarbiat Modares Univ, Sch Med Sci, Dept Med Genet, Tehran, Iran
[4] Monash Univ, Australian Regenerat Med Inst, Clayton, Vic 3800, Australia
关键词
Colorectal cancer; Cancer-associated fibroblasts; Exosome; Non-coding RNA; Angiogenesis; TUMOR MICROENVIRONMENT; CIRCULAR RNA; BIOMARKERS; HYPOXIA; GROWTH; GENES; PROLIFERATION; OPPORTUNITIES; MIGRATION;
D O I
10.1016/j.heliyon.2024.e31584
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Background: Circular RNAs (circRNAs) hold potential as diagnostic markers for colorectal cancer (CRC); however, their functional mechanisms remain incompletely elucidated. This work investigates the clinical implications of a unique set comprising six circRNAs derived from serum in CRC. Furthermore, we delve into the role of exosomal circ_0084043, originating from colorectal cancer -associated fibroblasts (CAFs), with a specific focus on its contribution to endothelial cell angiogenesis. Methods: The study analyzed circRNA levels in serum samples obtained from both CRC and control groups using qRT-PCR. Additionally, exosomes originating from colorectal CAFs and normal fibroblasts (NFs) were purified and confirmed by electron microscopy and Western blotting techniques. The proangiogenic effects of CAF -derived exosomal circ_0084043 were assessed in endothelial cells through proliferation, migration, and in vitro capillary tube formation assays. Gain- and loss -of -function experiments were employed to clarify the role of the circ_0084043/miR-140 -3p/HIF-1 alpha axis in endothelial cell angiogenesis, utilizing luciferase reporter assay, Western blotting, and ELISA for mechanism elucidation. Results: The candidate circRNAs (circ_0060745, circ_001569, circ_007142, circ_0084043, Circ_BANP, and CiRS-7) exhibited notably elevated expression in CRC patient sera compared to the levels observed in healthy individuals. Except for CiRS-7, all circRNAs showed elevated expression in CRC patients with positive lymph node metastasis and advanced tumor stages. Exosomes released by colorectal CAFs augmented endothelial cell proliferation, migration, and angiogenesis by upregulating VEGF expression and secretion. Circ_0084043 was highly detected in endothelial cells treated with CAF -derived exosomes. Silencing circ_0084043 reduced VEGFA expression and diminished CAF exosome-induced endothelial cell processes, indicating its pivotal role in angiogenesis. Circ_0084043 sponges miR-140 -3p, regulating HIF-1 alpha, and a reverse relationship was also identified between miR-140 -3p and VEGFA in endothelial cells. Inhibiting miR-140 -3p mitigated circ_0084043 knockdown effects in CAF exosome-treated endothelial cells. Co transfection of si-circ_0084043 and a miR-140-3p inhibitor reversed the inhibited migration and angiogenesis caused by circ_0084043 knockdown in CAF exosome-treated endothelial cells. Inhibiting miR-140-3p rescued reduced VEGFA expression due to circ_0084043 knockdown in endothelial cells exposed to CAF -derived exosomes, indicating modulation of the circ_0084043/ miR-140-3p/VEGF signaling in CAF -derived exosome-induced angiogenesis. Conclusions: This study unveiled a distinctive signature of six serum -derived circular RNAs, indicating their potential as promising diagnostic biomarkers for CRC. Importantly, exosomal circ_0084043 originating from colorectal CAFs was identified as playing a crucial role in endothelial cell angiogenesis, exerting its influence through the modulation of the miR-140-3p/HIF1 alpha/VEGF signaling axis.
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页数:16
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