Endothelial progenitor cell number and ERK phosphorylation serve as predictive and prognostic biomarkers in advanced hepatocellular carcinoma patients treated with sorafenib

Figures & data

Figure 1. Characterization of endothelial progenitor cells (EPCs) from PBMC. Viable cells were gated based on the expression of CD45_low CD146⁺, subsequently CD133⁺ cells were gated. Zebra plots showing reduction in ERK phosphorylation levels in EPCs of representative HCC patients after sorafenib/TACE treatment.

Figure 2. Effect of sorafenib treatment on ERK phosphorylation levels and absolute number of EPC. Pre-treatment samples of PBMC from HCC patients were treated with different concentrations of sorafenib (0–20 μM) in vitro and (A) the number of pERK⁺ EPC and (C) the absolute number of EPC were measured as described in Materials and Methods section. Each symbol (

) represents an individual HCC patient and lines represent mean values for the group. (B) The number of pERK⁺ EPC/mL and (D) absolute number of EPC in HCC patients before (▪) or after treatment (•) with sorafenib (in vivo) was measured as described in the materials and methods.

Figure 3. Effect of TACE on ERK phosphorylation levels and absolute number of EPC. Pre-treatment samples of PBMC from HCC patients (TACE) were treated with different concentrations of sorafenib (0–20 μM) in vitro and (A) the number of pERK⁺ EPC and (C) the absolute number of EPCs were measured as described in the Materials and Methods section. Each symbol (

) represents an individual HCC patient and lines represent mean values for the group. (B) The number of pERK⁺ EPC/mL and (D) the absolute number of EPCs in the PBMC of HCC patients before (▪) or after TACE (•) were measured as described in the materials and methods.

Figure 4. Effect of in vitro sorafenib treatment on ERK phosphorylation levels and absolute number of EPC. PBMC samples from HCC patients after treatment with sorafenib (A, C) or TACE (B, D) were treated with different concentrations of sorafenib (0–20 μM) in vitro and the number of pERK⁺ EPC and absolute number of EPC was measured as described in the Materials and Methods section. Each symbol (

) represents an individual HCC patient and lines represent mean values for the group.

Table 1. Clinical characteristics of patients.

	Sorafenib	TACE	ALL
	(n = 9)	(n = 8)	(n = 17)
Age
Median (Min,Max)	64 (53, 81)	69.5 (53, 84)	67 (53, 84)
Sex
Female	1 (11.1%)	1 (12.5%)	2 (11.8%)
Male	8 (88.9%)	7 (87.5%)	15 (88.2%)
Race
White	7 (77.8%)	7 (87.5%)	14 (82.4%)
Black	1 (11.1%)	0	1 (5.9%)
Am Indian, Aleutian, Eskimo	1 (11.1%)	0	1 (5.9%)
Asian Indian	0	1 (12.5%)	1 (5.9%)
Hepatitis B
No	9 (100%)	7 (87.5%)	16 (94.1%)
Yes	0	1 (12.5%)	1 (5.9%)
Hepatitis C
No	4 (44.4%)	4 (50%)	8 (47.1%)
Yes	5 (55.6%)	4 (50%)	9 (52.9%)
Alcohol Use
No	5 (55.6%)	6 (75%)	11 (64.7%)
Yes	4 (44.4%)	2 (25%)	6 (35.3%)
Cirrhosis
No	3 (33.3%)	4 (50%)	7 (41.2%)
Yes	6 (66.7%)	4 (50%)	10 (58.8%)
Previous Treatment
No	6 (66.7%)	6 (75%)	12 (70.6%)
Yes	3 (33.3%)	2 (25%)	5 (29.4%)
ECOG
0 = Normal, no complaints	6 (66.7%)	4 (50%)	10 (58.8%)
1 = Normal with effort	3 (33.3%)	4 (50%)	7 (41.2%)
AJCC Stage
1	2 (22.2%)	1 (12.5%)	3 (17.6%)
2	1 (11.1%)	3 (37.5%)	4 (23.5%)
3	6 (66.7%)	2 (25%)	8 (47.1%)
4A	0	2 (25%)	2 (11.8%)
BCLC Stage
B	8 (88.9%)	6 (75%)	14 (82.4%)
C	1 (11.1%)	2 (25%)	3 (17.6%)