Exosomes as potential diagnosis and treatment for liver cancer

Table 1 Potential biomarkers for liver cancer

Exosomal content	Sample	Expression	Isolation of exosomes	Content detection	Function	Ref.		Direction
HCC
Proteins
ANGPT2	Serum (n = 93)	Up	SBI	Immunoblotting and ELISA	Induces tumor angiogenesis	[14]	Potential targets
mRNAs
hnRNPH1	Serum (n = 223)	Up	Total exosome isolation reagent (Thermo Fisher Scientific Co.)	qRT-PCR	Associated with the Child–Pugh classification, portal vein tumor emboli, lymph node metastasis, TNM stage, and OS	[15]
LDH-C4	Serum (n = 212)	Up	exoRNeasy Serum/Plasma Midi Kit (Qiagen)	qRT-PCR	Related to treatments and recurrence prediction of HCC patients	[16]
miRNAs
miR-10b-5p	Serum (n = 37)	Up	Ultracentrifugation	qRT-PCR	Respectively, associated with early diagnosis and prognosis of HCC	[17]
miR-1247-3p	Serum (n = 135)	Up	Ultracentrifugation	qRT–PCR	Shows a positive correlation with lung metastasis in HCC patients	[18]	Potential targets
miR-125b	Serum (n = 218)	Up	SBI	qRT-PCR	Discriminate HCC patients with a high risk of recurrence and poor prognosis	[19]
miR-182	Serum and ascitic fluid	Up	exoRNeasy Serum/Plasma Midi Kit (Qiagen)	qRT-PCR	Up-regulated in NASH-induced liver cirrhosis with HCC compared to NASH-induced liver cirrhosis without HCC	[20]
miR-21	Serum (n = 79)	Up	SBI	qRT-PCR	Related to TNM stage and other prognostic factors	[21]
	Plasma (n = 150)	Up	SBI	qRT-PCR	Significantly higher in patients with HCC compared with cirrhotic patients and the control group	[22]
	Serum (n = 90)	Up	Total Exosome Isolation Reagent (Invitrogen)	qRT-PCR	Positively correlated with cirrhosis and tumor stage	[23]
	Serum (n = 95)	Up	Ultracentrifugation	qRT–PCR	Shows a positive correlation with survival in HCC patients	[24]	Potential targets
miR-215-5p	Serum (n = 37)	Up	Ultracentrifugation	qRT-PCR	Respectively, associated with early diagnosis and prognosis of HCC	[17]
miR-224	Serum (n = 139)	Up	Total Exosome Isolation Kit	qRT–PCR	Related to tumor size and differentiate HCC patients from healthy controls	[25]	Potential targets
miR23-a/b	Serum (n = 50)	Up	Ultracentrifugation	qRT–PCR	A promising target for future treatment of HCC	[26]	Potential targets
miR-301a	Serum and ascitic fluid (n = 52)	Up	exoRNeasy Serum/Plasma Midi Kit (Qiagen)	qRT-PCR	Up-regulated in NASH-induced liver cirrhosis with HCC compared to NASH-induced liver cirrhosis without HCC	[20]
miR-373	Serum and ascitic fluid (n = 52)	Up	exoRNeasy Serum/Plasma Midi Kit (Qiagen)	qRT-PCR	Up-regulated in NASH-induced liver cirrhosis with HCC compared to NASH-induced liver cirrhosis without HCC	[20]
miR-4661-5p	Serum (n = 720)	Up	SBI	qRT-PCR	Associated with the prognosis of patients with HCC	[27]
miR-638	Serum (n = 54)	Up	Ultracentrifugation	qRT–PCR	Promising for surveillance of HCC recurrence	[28]	Potential targets
miR-665	Serum (n = 40)	Up	SBI	qRT–PCR	Associated with tumor size, invasion, and clinical stage of HCC patients	[29]	Potential targets
miR-92a-3p	Plasma (n = 42)	Up	Ultracentrifugation	qRT–PCR	Shows a positive correlation with metastasis in HCC patients	[30]	Potential targets
miR-92b	Serum (n = 121)	Up	SBI	qRT-PCR	Prediction of posttransplant HCC early recurrence	[31]
miR-93	Serum (n = 108)	Up	Total Exosome Isolation Reagent (Invitrogen)	qRT–PCR	Correlated with stage, tumor size and predict patients' survival rate of HCC patients	[32]	Potential targets
miRNA-96	Plasma (n = 150)	Up	SBI	qRT-PCR	Significantly higher in patients with HCC compared with cirrhotic patients and the control group	[22]
lncRNAs
lncRNA-ATB	Serum (n = 79)	Up	SBI	qRT-PCR	Related to TNM stage and other prognostic factors	[21]
DANCR	Serum (n = 183)	Up	SBI	Digital droplet PCR (DDPCR)	Positively associated with HCV-HCC recurrence	[33]
lncRNA FAL1	Serum (n = 60)	Up	SBI	qRT-PCR	Play an oncogenic role in HCC	[34]	Potential targets
lnc-FAM72D-3	Serum (n = 180)	Up	Ultracentrifugation	qRT-PCR	Functions as an oncogene in HCC	[35]	Potential targets
lncRNA Jpx	Plasma (n = 100)	Up	SBI	qRT-PCR	Promising biomarkers for female patients with HCC	[36]
LINC00161	Serum (n = 112)	Up	Total Exosome Isolation Kit (Invitrogen)	qRT-PCR	A significant prediction of tumor growth and metastasis in HCC	[37]
	Serum (n = ?)	Up	-	qRT-PCR	Promote HCC tumorigenesis	[38]	Potential targets
lncRNA-RP11-583F2.2	Serum (n = 120)	Up	exoRNeasy Serum/Plasma Midi Kit (Qiagen)	qRT-PCR	Up-regulated in the serum of hepatocellular carcinoma patients as compared with hepatitis C virus patients and normal good health control	[39]
ENSG00000248932.1 ENST00000440688.1 ENST00000457302.2	Serum (n = 600)	Up	SBI	qRT-PCR	Potential fingerprints for the tumorigenesis prediction	[40]
circRNAs
circ_0070396	Plasma (n = 273)	Up	exoEasy Maxi Kit (QIAGEN)	qRT-PCR	Discriminate HCC individuals from patients with chronic hepatitis B and liver cirrhosis	[41]
circAKT3	Serum (n = 224)	Up	SBI	qRT-PCR	Associated with HCC recurrence and mortality	[42]
circ-DB	Plasma (n = 40)	Up	Ultracentrifugation	qRT-PCR	Promote the tumor growth	[43]	Potential targets
circPTGR1	Serum (n = 129)	Up	SBI	qRT-PCR	Promote HCC progression	[44]	Potential targets
circUHRF1	Serum (n = 643)	Up	SBI	qRT-PCR	Drive resistance to anti-PD1 immunotherapy	[45]	Potential targets
HB
miRNAs
miR-21	Serum (n = 64)	Up	SBI	qRT-PCR	Significantly higher in patients with HB	[46]

Up: Upregulated; SBI: Exo-Quick exosome precipitation solution; HCC: Hepatocellular carcinoma; HB: Hepatoblastoma; qRT-PCR: Quantitative reverse transcription polymerase chain reaction.

Table 2 Potential therapeutic drugs

Exosomal content	Sample	Expression	Isolation of exosomes	Content detection	Function	Ref.
HCC
miRNAs
miR-122	Serum (n = 75)	Down	SBI	qRT-PCR	Reflect the liver damage and residual liver function levels	[47]
	Plasma (n = 150)	Down	SBI	qRT-PCR	Significantly lower in patients with HCC compared with cirrhotic patients and the control group	[22]
miRNA-1298	Serum (n = 120)	Down	exoRNeasy Serum/Plasma MidiKit (Qiagen)	qRT-PCR	Down-regulated in patients of hepatocellular carcinoma compared with patients of hepatitis C virus and normal good health control	[39]
miR-320a	Serum (n = 209)	Down	SBI	qRT-PCR	Associated with lymph node metastasis, vein invasion, TNM stage, and survival of HCC patients	[48]
miR-320d	Serum (n = 150)	Down	Total Exosome Isolation Kit (Invitrogen)	qRT-PCR	Associated with clinicopathological parameters and prognosis of HCC patients	[49]
miR-638	Serum (n = 147)	Down	Total Exosome Isolation Kit (Invitrogen)	qRT-PCR	Influence liver carcinogenesis	[50]
miR-718	Serum (n = 59)	Down	Ultracentrifugation	qRT-PCR	Significantly different expression of HCC cases with recurrence after LT compared with those without recurrence	[51]
miR-744	Serum (n = 20)	Down	Ultracentrifugation	qRT–PCR	Facilitates the propagation and drug resistance of HCC cells	[52]
miR-9-3p	Serum (n = ?)	Down	Ultracentrifugation	qRT-PCR	A potential therapeutic target for HCC	[53]
lncRNAs
lnc-EPC1-4	Serum (n = 180)	Down	Ultracentrifugation	qRT-PCR	Function as a tumor suppressor gene	[35]
SENP3-EIF4A1	Serum (n = 6)	Down	SBI	qRT-PCR	Block HCC progression	[54]
circRNAs
circ-0051443	Plasma (n = 120)	Down	SBI	qRT-PCR	Suppress HCC progression	[55]
HB
miRNAs
miR-34s	Serum (n = 152)	Down	SBI	qRT-PCR	Significantly lower in patients with HB compared with the control group	[56]

Down: Downregulated; SBI: Exo-Quick exosome precipitation solution; HCC: Hepatocellular carcinoma; HB: Hepatoblastoma; qRT-PCR: Quantitative reverse transcription polymerase chain reaction.

Table 3 Potential therapeutic targets

Exosomal content	Sample	Expression	Content identification	Animal model (Yes/No)	Function	Ref.
HCC
Proteins
ENO1	Cancer cells-exosomes, tissue (n = 94)	Up	IHC staining	Y	Promotes HCC growth, metastasis, and further patient deterioration	[57]
miRNAs
miR-125a/b	TAMs-exosomes Tissue (n = 6)	Down	qRT-PCR	N	A possible therapeutic target in HCC	[58]
miR-150-3p	Fibroblasts-exosomes, tissues (n = 82)	Down	qRT–PCR	N	Abrogate HCC migration and invasiveness	[59]
miR-32-5p	Bel/5-FU-exosomes, tissue (n = 72)	Up	qRT–PCR	Y	Induce multidrug resistance in HCC	[60]
miR-320a	Cancer cells-exosomes, tissue (n = 6)	Down	qRT–PCR	Y	Mediates HCC tumor progression	[61]
miR-3682-3p	Cancer cells-exosomes, tissue (n = 8)	Down	qRT–PCR	Y	Attenuate angiogenesis and provides novel potential targets for liver cancer therapy	[62]
miR-378b	Cancer cells-exosomes, tissue (n = 105)	Up	qRT–PCR	Y	Enhance HCC cell progression and angiogenesis	[63]
lncRNAs
ASMTL-AS1	Cancer cells-exosomes, tissues (n = 70)	Up	qRT–PCR	Y	Aggravate the malignancy in residual HCC	[64]
PCED1B-AS1	Cancer cells-exosomes, tissues (n = 45)	Up	qRT–PCR	Y	Induce immunosuppression in HCC	[65]
circRNAs
circRNA Cdr1as	Cancer cells-exosomes, tissues (n = 42)	Up	qRT–PCR	Y	Promote the progression of HCC by sponging miR-1270 to upregulate AFP level	[66]

IHC: Immunohistochemistry; TAMs: Tumor-associated macrophages; Up: Upregulated; Down: Downregulated; HCC: Hepatocellular carcinoma; HB: Hepatoblastoma; qRT-PCR: Quantitative reverse transcription polymerase chain reaction.

Table 4 As a carrier for drug treatment

Drugs	Source of exosomes	Animal model (Yes/No)	Clinical sample (Yes/No)	Functions	Ref.
Norcantharidin	BMSCs-exosomes	Y	N	Induce cell cycle arrest, reduced tumor cell proliferation, increased apoptosis	[67]
siGRP78	BMSCs-exosomes	Y	N	Sensitize Sorafenib resistant cancer cells to Sorafenib	[68]
miR-214	hCEC-exosomes	N	Y (n = 6)	Enhances the anti-tumor efficacy of oxaliplatin and sorafenib on HCC cells	[69]

BMSCs: Bone marrow mesenchymal stem cell; hCEC: Human cerebral endothelial cell; HCC: Hepatocellular carcinoma.

Table 5 Exosomes from dendritic cells as potential therapeutic factors

Cargos	Source of exosomes	Animal model (Yes/No)	Clinical sample (Yes/No)	Functions	Ref.
Exosomes plus microwave ablation	DCs-exosomes	Y	N	Inhibit tumor growth and improve the immune microenvironment	[70]
Exosomes	DCs-exosomes	Y	N	Elicited strong antigen-specific immune responses and resulted in tumor growth retardation and prolonged survival rates in mice with ectopic	[71]

DCs: Dendritic cells.