Liquid biopsy in patients with hepatocellular carcinoma: Circulating tumor cells and cell-free nucleic acids

Table 2 Circulating cell-free DNA in hepatocellular carcinoma

Ref	HCC patients	Sample	Ethnicity	Background liver	Controls	cfDNA abnormalities	Target
						Methodolgy
Wong et al[105], 2000	25	Plasma/serum	Hong-Kong	HBV: 88%	55 (HD: 35, HV: 20)	Methylation	P16
	15	Buffy coat		HCV: 2%	35 (HD: 15, HV: 20)	MS-PCR
Wong et al[108], 2003	29/22	Plasma/serum	Hong Kong	NA	50 (HD and HV)	Methylation	P16INK4A
	29	Buffy coat			35 (HD: 15, HV: 20)	MS-PCR
Chu et al[109], 2004	46	Serum	Korea	HBV: 65%	23 (HD: 23)	Methylation	P16INK4A
				HCV: 22%		MS-PCR
Yeo et al[110], 2005	40	Plasma	Hong-Kong	HBV: 83%	10 (HV: 10)	Methylation	RASSF1A
						MS-PCR
Iizuka et al[96], 2006	52	Serum	Japan	HCV: 100%	46 (HD: 30, HV: 16)	Quantitative analysis	GSTP1
						Real-time PCR
Ren et al[97], 2006	79	Plasma	China	HBV: 85% LC: 86%	40 (HD: 20, HV: 20)	Quantitative analysis	NA
						Real-time PCR
						Allelic imbalance analysis	D8S258 and D8S264
Zhang et al[112], 2007	50	Serum	Taiwan	HBV: 22%	50	Methylation	P15, P16
				HCV: 16%	HV: 50	MS-PCR
Tan et al[111], 2007	8	Serum	Singapore	NA	72 (OT: 62, HV: 10)	Methylation	RUNX3
						MS-PCR
Chan et al[113], 2008	85	Serum	Hong-Kong	HBV: 92%	135 (HD: 63, HV: 72)	Methylation	RASSFIA
						RT-PCR
Chang et al[114], 2008	19	Plasma	China	HBV: 89%	17 (LC: 17)	Methylation	APC, GSTP1, RASSF1A, P16, E-cadherin
						MS-PCR
Iyer et al[115], 2010	28	Plasma	Egypt	HCV: 79%	NA	Methylation	APC, FHIT, P15, P16 and E-cadherin
				HNV: 18%		MS-PCR
Yang et al[98], 2011	60	Plasma	China	NA	50 (HD: 21, HV: 29)	Quantitative analysis	hTERT
						FQ-PCR
Szymañska et al[103], 2011	14	Plasma	China	Mostly HBV	NA	Single nucleotide mutation	R249S (TP53 mutation)
						SOMA
Iizuka et al[117], 2011	220	Serum	Japan	HCV: 100%	202 (HD: 202)	Methylation	SPINT2, SRD5A2
						MS-PCR
Huang et al[116], 2011	72	Plasma	China	HBV: 85%	37 (HD: 37)	Methylation	APC, GSTP1, RASSF1A, and SFRP1
						MSRE-qPCR
Huang et al[100], 2012	72	Plasma	China	HBV: 85%	115 (HD: 74, HV: 41)	Quantitative analysis	NA
						Real-time PCR
Chen et al[99], 2012	80	Serum	China	HBV: 100%	130 (HD: 80, HV: 50)	Quantitative analysis	NA
						Real-time PCR
Mohamed et al[118], 2012	40	Serum	Egypt	HCV: 100%	60 (HD: 40, HV: 20)	Methylation	RASSF1A
						Real-time PCR
Chen et al[101], 2013	39	Serum	China	HBV: 79%	45 (HV: 45)	Quantitative analysis	NA
						Real-time PCR
Piciocchi et al[102], 2013	66	Plasma	Italy	HCV: 51%	76 (HD: 76)	Quantitative analysis	hTERT
				Alcohol: 27%		Real-time PCR
Chan et al[95], 2013	4	Plasma	China	NA	20 (HD: 4, HV: 16)	Copy number variation	NA
						MPS
Sun et al[119], 2013	43	Serum	China	HBV: 86%	50 (HD: 24, HV: 26)	Methylation	TFPI2
						MS-PCR
Zhang et al[120], 2013	37	Serum	China	HBV: 100%	33 (HD: 33)	Methylation	DBX2, THY1
						Bead Chip, Hot-start PCR, Pyrosequencing
Huang et al[122], 2014	66	Serum	United States	HCV: 100%	43 (HD: 43)	Methylation	INK4A
				HCV and HBV: 6%		Pyrosequencing, MS-PCR
Han et al[121], 2014	160	Serum	China	HBV: 22%	133 (HD: 88, HV: 45)	Methylation	TRG5
						MS-PCR
Ji et al[123], 2014	121	Serum	China	HBV: 83%	68 (HD: 37, HV: 31)	Methylation	MT1M
						MS-PCR
Kuo et al[124], 2014	40	Plasma	Taiwan	NA	34	Methylation	HOXA9
						MS-PCR
Jiang et al[104], 2015	90	Plasma	Hong-Kong	NA	135 (HD: 103, HV: 32)	Copy number variation	NA
						CAZA

NA: Not applicable; HD: Hepatic disease without evidence of HCC; HV: Healthy volunteers; RT-PCR: Reverse transcriptase polymerase chain reaction; SOMA: Short oligonucleotide mass analysis; CAZA: Chromosome arm-level z-score analysis; MPS: Massively parallel sequencing; MS-PCR: Methylation-specific PCR; RT-PCR: Real time polymerase chain reaction; MSRE-qPCR: Methylation-sensitive restriction enzymes-based quantitative PCR; FQ-PCR: Real-time quantitative fluorescent polymerase chain reaction.