Comparison of the disease-free survivals among patients with various risk scores, which were defined as the number of independent predictors (positive HURP expression in HCC cells, AST ≥50 U/L, cytological grade ≥3, and tumor number >1) carried by each patient.

<p>(A) Comparison of the disease-free survivals among patients with various risk scores ranged from 0 to 3. (B) The patients with risk score = 0 and those with risk score = 1 were merged.</p

Association between clinical parameters and disease-free survival.

<p>Association between clinical parameters and disease-free survival.</p

Basic clinical parameters for HCC patients.

a<p>Other patients had tumors removed surgically.</p>b<p>Post-surgery specimens.</p

Comparison of the disease-free survivals between HCC patients with and without a statistically significant clinicopathological feature.

<p>n, number of HCC patients at risk.</p

Independent predictors of disease-free survival in the stepwise multivariate Cox proportional hazard model.

<p>Independent predictors of disease-free survival in the stepwise multivariate Cox proportional hazard model.</p

Immunohistochemistry analysis for HURP expression in aspirated human HCC cells in 8 representative cases.

<p>HURP was stained in brown color.</p

DataSheet_1_WWOX-rs13338697 genotype predicts therapeutic efficacy of ADI-PEG 20 for patients with advanced hepatocellular carcinoma.docx

BackgroundPrevious studies have identified three single nucleotide polymorphisms (SNPs): GALNT14-rs9679162, WWOX-rs13338697 and rs6025211. Their genotypes are associated with therapeutic outcomes in hepatocellular carcinoma (HCC). Herein, we examined whether these SNP genotypes could predict the clinical outcome of HCC patients treated with ADI-PEG 20.MethodsTotally 160 patients with advanced HCC, who had previously been enrolled in clinical trials, including 113 receiving ADI-PEG 20 monotherapy (cohort-1) and 47 receiving FOLFOX/ADI-PEG 20 combination treatment (cohort-2), were included retrospectively.ResultsThe WWOX-rs13338697-GG genotype was associated with favorable overall survival in cohort-1 patients (P = 0.025), whereas the rs6025211-TT genotype was associated with unfavorable time-to-tumor progression in cohort-1 (P = 0.021) and cohort-1 plus 2 patients (P = 0.008). As ADI-PEG 20 can reduce plasma arginine levels, we examined its pretreatment levels in relation to the WWOX-rs13338697 genotypes. Pretreatment plasma arginine levels were found to be significantly higher in patients carrying the WWOX-rs13338697-GG genotype (P = 0.006). We next examined the association of the WWOX-rs13338697 genotypes with WWOX tissue protein levels in 214 paired (cancerous/noncancerous) surgically resected HCC tissues (cohort-3). The WWOX-rs13338697-GG genotype was associated with decreased tissue levels of WWOX and ASS1. Mechanistic studies showed that WWOX and ASS1 levels were downregulated in hypoxic HCC cells. Silencing WWOX to mimic low WWOX protein expression in HCC in patients with the WWOX-rs13338697-GG genotype, enhanced HIF1A increment under hypoxia, further decreased ASS1, and increased cell susceptibility to ADI-PEG 20.ComclusionIn summary, the WWOX-rs13338697 and rs6025211 genotypes predicted treatment outcomes in ADI-PEG 20-treated advanced HCC patients. The WWOX-rs13338697-GG genotype was associated with lower tissue WWOX and ASS1 levels and higher pretreatment plasma arginine levels, resembling an arginine auxotrophic phenotype requires excessive extracellular arginine supply. Silencing WWOX to mimic HCC with the WWOX-rs13338697-GG genotype further stimulated HCC cell response to hypoxia through increased HIF1A expression, leading to further reduction of ASS1 and thus increased cell susceptibility to ADI-PEG 20.</p

Basic clinical data of patients included.

<p>a Comparison between patients with tumor number = 1 and those with tumor number >1.</p><p>b Median (range).</p><p>c Mann-Whitney test.</p

Comparison of virological factors between occult and overt HBVCI patients with HCV-related HCC.

<p>a Median (range); b Mann-Whitney test; c Kaplan-Meier analysis; d Estimation is limited to the largest survival time if it is censored.</p><p>After Bonferroni correction for the alpha error, P<0.006 was considered statisitical significant.</p><p>Lower detection limit of intrahepatic HBV-DNA, 120 IU/g; Lower detection limit of intrahepatic HCV-RNA, 1500 IU/g.</p

Association between ephrinA5 isoforms, clinical parameters and disease-free survival/overall survival.

a<p>Kaplan-Meier analysis with log rank test.</p>b<p>Relative expression of ephrinA5 mRNA assessed by real-time RT-PCR using peritumoral liver tissues.</p><p>CI: Confidence Interval,</p>*<p>P<0.05.</p