Function and biomedical implications of exosomal microRNAs delivered by parenchymal and nonparenchymal cells in hepatocellular carcinoma

Table 1 Function of exosomal microRNAs from interstitial cells in the liver

miRNA species in exosomes	Exosome secreting cells	Exosome isolation methods	Target cells	miRNA expression of exosome	Downstream targets	Functions of miRNA	Additional information	Ref.	Year
miR-148a-3p	Primary fibroblasts (the HSC cell line LX-2)	The ExoQuick-TC kit	Human HCC cell lines PLC, HCCLM3, and SMMC-7721	Reduced in the exosomes of HSCs after cocultivation with primary liver cancer-associated fibroblasts	ITGA5/PI3K/Akt axis	Inhibited HCC cell malignancy	Primary fibroblasts were isolated from primary HCC tumor and paired peritumor tissues in 17 primary HCC patient samples	[78]	2022
miR-335-5p	The HSC cell line LX-2	Ultracentrifugation	Human HCC cell lines MHCC97H, MHCC97L, HepG2, and Huh7	Reduced in the exosomes of fibroblasts as well as in HCC cells after cocultivation	CDC42? CDK2?	Inhibited neighboring cancer cell proliferation, invasion, and motility	-	[79]	2019
miR-320a	CAFs	Life Technology exosome precipitation solution	Human HCC cell lines MHCC97-H, SMMC-7721, Huh7, and the human normal liver cell line 7702	Reduced in the exosomes of CAFs derived from human HCC patients	PBX3	Inhibited HCC cell proliferation and metastasis ability	PAFs and CAFs derived from 6 pairs of matched primary hepatocarcinoma and adjacent tumor-free tissues (5 cm from the cut edge of the tumor edge)	[87]	2017
miR-150-3p	CAFs	0.22-µm PVDF filter and Total Exosome Isolation Reagent	Human HCC cell lines Huh7 and Hep3B	Decreased in CAF-derived exosomes	-	Inhibited HCC proliferation and metastasis	Stromal fibroblasts isolated from tumor tissue and adjacent (> 5 cm from the tumor edge) tissues from 6 HCC patients	[88]	2021
miR-20a-5p	CAFs	Centrifuged and filtered through a 0.22-µm PVDF membrane	Human HCC cell lines SMMC7721, Huh7, YY8103, Hep3B, Focus, HepG2, and HCCLM3 and a normal liver cell line MIHA, WRL68	Higher in exosomes from cancer tissues than in matched adjacent para-tumoral tissues	LIMA1	Contributed to HCC cell proliferation, metastasis, and EMT	CAFs were from the HCC tissues and NFs in paired adjacent normal tissues from 92 HCC patients	[89]	2022
miR-214	hCECs	Centrifuged and filtered through a 0.22-µm PVDF membrane and ultracentrifugation	Human HCC cell lines HepG2, Hep3B, the human liver epithelial cell line THLE-2	Lower levels in HCC cells than in normal human liver epithelial cells	P-gp/SF3B3	Reduced cancer cell viability and invasion compared with monotherapy with oxaliplatin or sorafenib	-	[103]	2021
miR-23a/b	Adipose cell mouse preadipocyte 3T3-L1 cells	Differential centrifugation	The human HCC cell lines BEL-7402 and BEL-7402/5-Fu murine hepatoma cell line Hepa1-6	High in exosomes from HCC patients with high BFR	VHL/HIF-1α	Promoted HCC cell growth and migration	Adipose cells were isolated from human tumor tissues from obese and nonobese patients	[95]	2019
miR-142, miR-223	Monocyte-derived macrophages; human acute monocytic leukemia THP-1, B-lymphoblastoid 721.221, and murine lymphoblast-like mastocytoma P815 cell lines	Microfiltration and ultracentrifugation	The human HCC cell lines Huh7 and HepG2	High when cocultured with HCC cells	STMN-1	Inhibited HCC proliferation	PBMCs were isolated from lymphocyte cones or fresh blood by density gradient centrifugation and were incubated for 2 h in plastic plates before the flask was washed intensively to remove any nonadherent cells. After 4 d of incubation in serum-free medium supplemented with 1% autologous serum, adherent cells were washed with PBS and cultured in standard DMEM-based medium for 3-6 extra days to generate monocyte-derived macrophages phenotyped to be CD14+, CD11a+, CD3−, CD56−, and CD19−	[119]	2013
miR-490	Human MC line HMC-1 (treated with HCV-E2)	Total exosome separation reagent from Invitrogen	The human HCC cell lines HepG2 and HepG3b	High when HCV-E2-stimulated MC-derived exosomes were incubated with the two types of HCC cells for 24 h compared with the incubation with normal MC-derived exosomes	ERK1/2	Inhibited HCC proliferation		[121]	2017
miR-223	Human NK cell line NK92-MI	Differential centrifugation	The human HSC line LX-2	Higher in exosomes derived from NK cells than in parental NK-92MI cells	AGT7	Attenuated TGF-β1-induced HSC activation and inhibited liver fibrosis	LX-2 cells were treated with TGF-β1 (5 ng/mL) for 24 h to stimulate HSC activation. LX-2 cells in the exosomes derived from NK cells-treated groups were pretreated with exosomes derived from NK cells (10 μg/mL) before TGF-β1 treatment. LX-2 cells in the rapamycin-treated groups were pretreated with the autophagy activator rapamycin (2 mM) in DMSO for 12 h before TGF-β1 treatment	[120]	2020
miR-125a/b	TAMs	ExoQuick exosome precipitation solution	The human HCC cell lines Huh7, HepG2, and BEL-7404	Downregulated in exosomes from HCC-associated macrophages	CD90	Suppressed HCC cell growth and sphere formation	TAMs and nontumor macrophages were isolated from primary human HCC, adjacent nontumor liver tissues from 6 patients with HCC	[115]	2019
miR-628-5p	M1 macrophage	-	The human HCC cell lines Huh7, HCCLM3, Hep3B, and MHCC97H, immortalized human liver epithelial THLE-3 cell line	High in M1-exosomes	METTL14/circFUT4/CHMP14B	Inhibited HCC cell progression	THP-1 cells were differentiated into M0 macrophages by a 24 h incubation with 150 nM phorbol 12-myristate 13-acetate followed by a 24 h incubation in RPMI medium. M0 macrophages were polarized to M1 macrophages by incubation with 20 ng/mL IFN-γ and 10 pg/mL lipopolysaccharide	[118]	2022
miR-92a-2-5p	M2 macrophage (monocytic leukemia cell line THP-1)	Centrifuged and filtered through a 0.22-µm PVDF membrane and ultracentrifugation	Human liver cancer SK-HEP-1 and HepG2 cell lines, HA22T cell line, and murine HCC Hepa 1-6 cell line	Increased after coculture with liver cancer cells	AR/PHLPP/p-AKT/β-catenin signaling	Promoted HCC growth and invasiveness	To induce differentiation into macrophages, THP-1 cells were cultured with 100 ng/mL PMA (Sigma) for 48 h, and the macrophage was cultured with the addition of DMSO to promote M2 polarization	[112]	2020
miR-660-5p	M2 macrophage (monocytic leukemia cell line THP-1)	Differential centrifugation	Human HCC cell lines HepG2 and Bel-7402	High	KLF3	Augmented the tumorigenic ability of HCC cells	THP-1 monocytes were stimulated by 100 ng of phorbol 12-myristate 13-acetate (Sigma-Aldrich, MO, United States) for 48 h, thus differentiating into M0 macrophages. Then, M0 macrophages were treated with 20 ng/mL interleukin 4 (AF-200–04-5, PeproTech, NJ, United States) for 72 h to polarize into M2 macrophages	[114]	2021
miR-27a-3p	M2 macrophage (monocytic leukemia cell line THP-1)	SBI ExoQuick-TC Kit	Human HCC cell lines Huh7, 97H, HepG2, LM3, and SMMC-7721	-	TXNIP	Induced the cancer stemness of HCC	Differentiation of THP-1 cells to macrophages was performed using 200 ng/mL phorbol myristic acetate, and the cells were then cultured with 20 ng/mL interleukin-4 for 72 h to induce M2-type polarization	[113]	2021
miR-142-3p	TAMs treated by propofol (the murine macrophage cell line Raw 264.7 cells)	Differential centrifugation	The murine HCC cell line Hepa1-6	Dose-dependent increase when treated with propofol	RAC1	Enhanced the antitumor activity of propofol	Raw 264.7 cells were cultured in complete RPMI 1640 with 10% FBS and treated with propofol (dissolved in RPMI 1640) in complete medium. TAMs were isolated from tumor-bearing mice treated with 0 mg/kg, 20 mg/kg, and 50 mg/kg propofol by i.p. injection	[117]	2014
miR-375	TAMs (IL-2 induced)	Total exosome isolation reagent	The human HCC cell lines HepG2 and QJY–7703	High	-	Ameliorated HCC development and progression	Primary human HCC specimens were collected from patients who suffered from hepatectomy. The macrophages were isolated and cultured by Percoll (GE Healthcare) density gradient centrifugation. TAMs were treated with IL-2 for 24 h before the supernatants were collected. The treatment concentration was 20 ng/mL	[116]	2022

AGT7: Autophagy-related 7; AR: Androgen receptor; BFR: Body fat ratio; CAFs: Cancer-associated fibroblasts; CDC42: Cell division cycle 42; CDK2: Cyclin dependent kinase 2; EMT: Epithelial-mesenchymal transition; ERK1/2: Extracellular regulated protein kinases 1/2; Exos: Exosomes; HCC: Hepatocellular carcinoma; hCECs: Human cerebral endothelial cells; HCV-E2: Hepatitis C virus E2 envelope glycoprotein; HIF-1α: Hypoxia-inducible factor 1α; HSCs: Hepatic stellate cells; i.p.: Intraperitoneal; ITGA5: Integrin α5; KLF3: Kruppel-like factor 3; LIMA1: LIM domain and actin binding 1; MCs: Mast cells; METTL14: Methyltransferase-like 14; miR: MicroRNA; NK: Natural killer; PAFs: Para-neoplastic fibroblasts; PBMCs: Peripheral blood mononuclear cells; PBX3: Pre-B-cell leukemia homeobox 3; P-gp: P-glycoprotein; PI3K: Phosphoinositide 3-kinase; RAC1: Rac family small GTPase 1; SF3B3: Splicing factor 3b subunit 3; STMN1: Stathmin-1; TAMs: Tumor-associated macrophages; TXNIP: thioredoxin-interacting protein; VHL: Von Hippel-Lindau.