MiR-21 in Substance P-induced exosomes promotes cell proliferation and migration in human colonic epithelial cells

• Published in: American journal of physiology: Gastrointestinal and liver physiology 2019-12, Vol.317 (6), p.G802-G810
• Main Authors: Bakirtzi, Kyriaki, Man Law, Ivy Ka, Fang, Kai, Iliopoulos, Dimitrios, Pothoulakis, Charalabos
• Format: Article
• Language: English
• Subjects: Animals; Biosynthesis; Cell culture; Cell growth; Cell migration; Cell Movement - immunology; Cell proliferation; Cell Proliferation - physiology; Cell signaling; Cells, Cultured; Colitis - metabolism; Colon; Colon - metabolism; Colon - pathology; Culture media; Epithelial cells; Exosomes; Exosomes - metabolism; Flow Cytometry; Fluorescence microscopy; Humans; Inflammation - metabolism; Intestinal Mucosa - metabolism; Mice; MicroRNAs - metabolism; miRNA; Polymerase chain reaction; Receptors, Neurokinin-1 - metabolism; Signal Transduction; Substance P; Substance P - metabolism
• ISSN: 0193-1857; 1522-1547; 1522-1547
• DOI: 10.1152/ajpgi.00043.2019

Exosomes are cellular vesicles involved in intercellular communication via their specialized molecular cargo, such as miRNAs. Substance P (SP), a neuropeptide/hormone, and its high-affinity receptor, NK-1R, are highly expressed during colonic inflammation. Our previous studies show that SP/NK-1R signaling stimulates differential miRNA expression and promotes colonic epithelial cell proliferation. In this study, we examined whether SP/NK-1R signaling regulates exosome biogenesis and exosome-miRNA cargo sorting. Moreover, we examined the role of SP/NK-1R signaling in exosome-regulated cell proliferation and migration. Exosomes produced by human colonic NCM460 epithelial cells overexpressing NK-1R (NCM460-NK1R) were isolated from culture media. Exosome abundance and uptake were assessed by Western blot analysis (abundance) and Exo-Green fluorescence microscopy (abundance and uptake). Cargo-miRNA levels were assessed by RT-PCR. Cell proliferation and migration were assessed using xCELLigence technology. Colonic epithelial exosomes were isolated from mice pretreated with SP for 3 days. Cell proliferation in vivo was assessed by -67 staining. SP/NK-1R signaling in human colonic epithelial cells (in vitro) and mouse colons (in vivo) increased ) exosome production, ) the level of fluorescence in NCM460s treated with Exo-Green-labeled exosomes, and ) the level of miR-21 in exosome cargo. Moreover, our results showed that SP/NK-1R-induced cell proliferation and migration are at least in part dependent on intercellular communication via exosomal miR-21 in vitro and in vivo. Our results demonstrate that SP/NK-1R signaling regulates exosome biogenesis and induces its miR-21 cargo sorting. Moreover, exosomal miR-21 promotes proliferation and migration of target cells. Substance P signaling regulates exosome production in human colonic epithelial cells and colonic crypts in wild-type mice. MiR-21 is selectively sorted into exosomes induced by Substance P stimulation and promotes cell proliferation and migration in human colonocytes and mouse colonic crypts.