Expression Of Glypican-3 In Hepatocellular Carcinoma And Its Association With Postoperative Early Recurrence

1. Objective:To indentify the independent risk factors for ER of HCC after curative resection.To evaluate the usefulness of GPC3 as a risk factor for early intrahepatic recurrence of HCC after hepatic resection.2. Methods:A total fifty-seven patients who were performed curative hepatic resection for HCC were enrolled and divided into two groups:early recurrence(ER) group and no early recurrence (NER) group. ER group had the time interval from hepatic resection to recurrence within 1 year, while no early recurrence (NER) group had not intrahepatic recurrence in 2 year after hepatic resection.11 general clinical parameters (gender:male/femal; age;HbsAg:+/-;HbeAg:+/-;HCV:+/-;HBV DNA: normal/abnormal; Child class:A/B;AFP:< 400μg/l/≥400μg/l; ALT:< 80U/L/≥80U/L;liver cirrhosis:+/-; perioperative blood transfusion:+/-) and and 8 tumor parameters(tumor size:<5cm/≥5cm;No.of tumor:sigle/multiple; satellite nodular:+/-;lymphoid node metastasis:+/-; microvascular invasion:+/-;portal vein invasion:+/-;capsule invasion:+/-;Edmonson grade:Ⅰ-Ⅱ/Ⅲ-Ⅳ) were compared between 2 groups using univariate and multivariate analysis.Our earlier stage result of cDNA microarrays showed that the expression of GPC3 in ER group was almost 4 times higher than in NER group. Quantitative real-time RT-PCR and Immunohistochemistry was used to assess GPC3 expression.χ2 test, the Fisher exact test or Student t test were used for univariate analysis and logistic regression analysis for multivariate analysis. All statistics were performed by SPSS (Version 13.0) software and P<0.05 was considered to be statistically significant 3. Results:(1) Risk factors for early intrahepatic recurrence by univariate analysisResults of univariate analysis indicated that liver cirrhosis(P=0.022),tumor size(> 5cm)(P=0.000), microvascular invasion (P=0.022) and ALT(>80U/L) (P=0.003)were significantly different between 2 groups.These risk factors may be associated with early intrahepatic recurrence.(2) Expression of GPC3 mRNA in HCC by quantitative real-time RT-PCRResults of real time RT-PCR revealed that the average GPC3 expression in ER group (3.977±0.676) was higher than NER group (3.503±0.926)(t=2.213, P=0.031).(3) Expression of GPC3 protein in HCC by immunohistochemistryImmunohistochemistry indicated normal liver tissue sections were negative for GPC3.Of all 57 cases,68.42%(39/57) section showed the reactivity of GPC3. 82.8%(24/29) sections in ER group were positive for GPC3,while 53.6%(15/28)in NER group (χ2=5.617, P<0.05). The positive expression rate of GPC3 protein in HCC tissues had no relation with general clinical factors and tumor factors (p> 0.05).(4) Independent risk factors for early intrahepatic recurrenceLogistic regression analysis indicated that GPC3-positive(OR:7.110;95%CI: 1.158-43.639), tumor size(OR:8.170;95%CⅠ:1.626-41.036), microvascular invasion (OR:10.022;95%CⅠ:1.141-88.040)were independent risk factors for early intrahepatic recurrence.4. Conclusion:(1) GPC3-positive,tumor size(>5cm),microvascular invasion were independent risk factors for early intrahepatic recurrence. (2) The positive expression rate of GPC3 protein in HCC tissues had no relation with general clinical factors and tumor factors. GPC3 was confirmed be associated with early recurrence of HCC after curative resection.