Decreased exosome-delivered miR-486-5p is responsible for the peritoneal metastasis of gastric cancer cells by promoting EMT progress

Abstract

Background The present study aims to investigate the preliminary mechanism underlying the peritoneal metastasis of gastric cancer cells. Methods Exosomes from GC9811 cells (Con-Exo) and from GC9811-P cells (PM-Exo) were extracted by ultracentrifugation, which were identified with transmission electron microscopy (TEM) and nanoparticle trafficking analysis, as well as the expression of CD9, CD63, and CD81 detected by Western blot assay. α-SMA expression was determined by immunofluorescence assay and Western blot assay. The levels of Snail1, E-cadherin, and Actin-related protein 3 (ACTR3) were evaluated by Western blot assay. MiRNA array was performed on exosomes to screen the differentially expressed miRNAs. The expressions of miRNAs, SMAD2, CDK4, and ACTR3 were determined by QRT-PCR. The delivery of miR-486-5p was confirmed by laser confocal detection. Results Firstly, TEM, nanoparticle trafficking analysis, and Western blot assays were used to confirm the successful extraction of Con-Exo and PM-Exo. The incubation of Con-Exo and PM-Exo could decrease E-cadherin expression and increase of α-SMA respectively in HMrSV5 cells, with the increased proportion of fusiform cells. More significant changes were observed in PM-Exo-treated HMrSV5 cells. Secondary, compared to Con-Exo, miR-486-5p and miR-132-3p were found downregulated, and miR-132-5p was found upregulated in PM-Exo. The transfection of miR-486-5p and miR-132-3p was observed to suppress EMT, and the transfection of miR-132-3p was observed to induce EMT. Laser confocal detection confirmed the delivery of miR-486-5p from gastric cancer cells to HMrSV5 cells through exosomes. Lastly, the expression of Mothers against decapentaplegic homolog 2 (SMAD2), cyclin-dependent kinase 4 (CDK4), and ACTR3 was found to be downregulated via miR-486-5p. Conclusion Decreased delivery of miR-486-5p via exosomes might be responsible for the peritoneal metastasis of gastric cancer cells by promoting epithelial-mesenchymal transition progress.

Alpha-hederin reprograms multi-miRNAs activity and overcome small extracellular vesicles-mediated paclitaxel resistance in NSCLC

“…We hypothesized that α-hederin might regulate A549T-derived sEVs cargos involved in TGFβ/SMADs and SREBF1/FASN signaling. A variety of non-coding RNAs, including microRNAs (miR-21-5p, miR-23a-3p, miR-125b-5p, miR-145-5p, miR-26a-5p, miR-6766-3p, miR-671-5p, let-7a-5p, miR-486-5p, miR-29c-3p, and miR-29c-5p) and long non-coding RNAs (lncRNA ZEB1-AS1, LINC00960, LINC02470, lncRNA SOX2-OT, and lncRNA CDKN2B-AS1), were shown to be sorted into sEVs or exosomes and directly targeted regulatory genes in the TGFβ/SMADs pathway ( Solé et al, 2015 ; Fang et al, 2016 ; Hu et al, 2019a ; Huang et al, 2020 ; Wang et al, 2021b ; Chen et al, 2021 ; Wang et al, 2021c ; Lin et al, 2021 ; Wu et al, 2021 ; Zhou et al, 2021 ; Wu et al, 2022a ; Luo et al, 2022 ). Among these non-coding RNAs, we observed that miR-21-5p, miR-23a-3p, and miR-125b-5p decreased after treatment with A549T-derived sEVs and increased after further treatment with α-hederin ( Figure 6A ).…”

The SMAD2/miR-4256/HDAC5/p16INK4a signaling axis contributes to gastric cancer progression

“…Liao et al confirmed that HDAC5 was highly expressed in GC tumor tissues and promoted the proliferation and migration capability of GC cells [ 30 ]. Moreover, Lin et al showed that HDAC5 was upregulated in GC cells and boosted the invasiveness of GC cells by enhancing matrix metalloproteinase 9 (MMP9) [ 31 ]. MMP9 has been confirmed as an important gene implicated in tumor invasion and metastasis through epithelial-mesenchymal transition (EMT) processes [ 32 ].…”

The biological role of extracellular vesicles in gastric cancer metastasis

“…When GC cells were co-cultured with EVs rich in GKN1, the expression of E-cadherin in GC cells increased, while the expression of proteins such as N-cadherin significantly decreased, suggesting that GKN1 derived from EVs inhibited GC metastasis by suppressing EMT ( Yoon et al, 2020 ). Studies have also shown that biologically active substances such as miR-486-5p and circ-ITCH delivered by EVs could inhibit the metastasis of GC by suppressing EMT ( Wang et al, 2021b ; Lin et al, 2021 ).…”