Prognostic Value of ALT, AST, and AAR in Hepatocellular Carcinoma with B-Type Hepatitis-Associated Cirrhosis after Radical Hepatectomy.

BACKGROUND: Alanine aminotransferase (ALT), aspartate aminotransferase (AST), and AST to ALT ratio (AAR) were shown to be associated with prognosis in some groups of hepatocellular carcinoma (HCC). However, their clinicopathologic and prognostic roles in HCC patients with B-type hepatitis-associated cirrhosis (HBAC) have not been comprehensively investigated. The present study aimed to address the issues. METHODS: A total of 125 patients with HCC and HBAC after radical hepatectomy were included. The correlations of ALT, AST, and AAR with clinicopathologic parameters, overall/recurrence-free survival, overall/early recurrence, and post-recurrence survival were evaluated using univariate and multivariate analyses. RESULTS: ALT and AST, which positively correlated with each other, had significant relationships with tumornode-metastasis (TNM) stage and Edmondson-Steiner grade. In univariate analyses, ALT and AST were predictive for early recurrence, overall and recurrence-free survival, while ALT and AST was associated with overall recurrence and post-recurrence survival, respectively. However, only AST was marginally significant in multivariate tests for early recurrence and post-recurrence survival. As for AAR, no significant prognostic relevance was found. CONCLUSIONS: Our data suggest that ALT and AST, but not AAR, might be potential predictors of post-resectional outcome in HCC with HBAC. These effects might depend on their associations with crucial clinicopathologic variables.

Early recurrence in large hepatocellular carcinoma after curative hepatic resection: prognostic significance and risk factors.

BACKGROUND/AIMS: So far, prognostic significance and risk factors of early recurrence after curative resection in large hepatocellular carcinoma (LHCC) remain unclear. The present study aimed to answer these questions. METHODOLOGY: Clinical, pathologic and follow-up data of consecutive 116 patients with LHCC (>5cm) after curative resection were collected and analyzed. The recurrence pattern of LHCC was also compared with that of 55 patients with small HCC (SHCC, ≤5cm). RESULTS: Forty-five patients (38.8%) with LHCC developed recurrence within 1 year after surgery (defined as early recurrence), with a significantly higher ratio contrast to those with SHCC. Univariate analysis showed that age, HBsAg positivity, satellite nodule, TNM stage, resection extent and early recurrence served as significant indicators of post-recurrence overall survival in recurrent LHCC. In addition, only early recurrence was proven to be significant in multivariate Cox regression test. On the other hand, age, HBsAg positivity, portal vein tumor thrombosis, microvascular invasion, TNM stage, Edmondson-Steiner grade and resection extentwere related to early recurrence in LHCC. Among them, microvascular invasion and Edmondson-Steiner grade were independent predictors. CONCLUSIONS: Patients with early recurrence carried very poor post-recurrence prognosis in LHCC. The microvascular involvement and differentiation grade might be particularly helpful for prediction of early recurrence.

Risk factors of microvascular invasion, portal vein tumor thrombosis and poor post-resectional survival in HBV-related hepatocellular carcinoma.

BACKGROUND/AIMS: Microvascular invasion (MVI) and portal vein tumor thrombosis (PVTT) associated factors in hepatocellular carcinoma (HCC) were previously shown. However, those for HBV-related HCC remain unknown. This study aimed to investigate the risk factors of MVI, PVTT and poor prognosis in this type of HCC. METHODOLOGY: Consecutive 130 patients with HBV-related HCC were enrolled. The impact of variables on MVI, PVTT and post-resectional survival was determined by uni- and multi-variate analyses. RESULTS: By Chi-square analysis, Edmondson-Steiner grade and tumor size were related to MVI, whereas Edmondson-Steiner grade and MVI were associated with PVTT. Logistic regression identified Edmondson-Steiner grade as the sole independent determinant of MVI, but none is significant for PVTT. Tumor size carried high sensitivity and negative predictive value in prediction of MVI. Survival estimation revealed that Edmondson-Steiner grade, tumor size, PVTT, MVI, satellite nodule, TNM stage and AFP level were prognostic for overall and disease-free survival, but only Edmondson-Steiner grade was of independent implication for both. Besides, tumor size independently predicted overall survival. CONCLUSIONS: In HBV-related HCC, differentiation degree might play an important role in vascular involvement, tumor size might be helpful in primary screening of MVI, differentiation and tumor size might be particularly significant as prognostic markers.

Risk factors of poor prognosis and portal vein tumor thrombosis after curative resection of solitary hepatocellular carcinoma.

BACKGROUND: Predictors of poor prognosis of solitary hepatocellular carcinoma (SHCC), a subgroup encompassing most patients with the malignancy, are still controversial. Hence, risk factors for portal vein tumor thrombosis (PVTT) in SHCC are obscure. The present study was designed to address this issue. METHOD: Clinicopathological and follow-up data for 156 consecutive patients with SHCC following curative hepatic resection were analyzed using uni- and multi-variate analyses. RESULTS: Univariate analysis showed that PVTT, tumor-node-metastasis (TNM) stage, Edmondson-Steiner grade and preoperative serum alpha-fetoprotein (AFP) level were associated with the overall and disease-free survival, whereas tumor size only influenced the overall survival. In multivariate Cox regression tests, Edmondson-Steiner grade and TNM stage were independent prognostic markers for both overall and disease-free survival. In addition, the Chi-square test showed that AFP level and Edmondson-Steiner grade were correlated with PVTT. Among them, only Edmondson-Steiner grade was shown to be of independent significance for PVTT in multi-variate logistic regression analysis. Additionally, AFP, the sole preoperative factor for PVTT, was not adequately sensitive and specific. CONCLUSIONS: Factors relating to post-surgical prognosis and PVTT in SHCC are all tumor-related. Of these, Edmondson-Steiner grade and TNM stage might be of particular importance in survival analysis. In addition, accurate prediction of PVTT by clinicopathological parameters before surgery remains difficult.

The significance of serum AFP cut-off values, 20 and 400 ng/mL in curatively resected patients with hepatocellular carcinoma and cirrhosis might be of difference.

BACKGROUND/AIMS: The significance of preoperative serum a-fetoprotein (AFP) level in hepatocellular carcinoma (HCC) treated with different modalities remains controversial. Besides, many cut-off values have been used. The present study aims to clarify significance of two major ones in HCC with cirrhosis. METHODOLOGY: One hundred and thirty eight consecutive cirrhotic patients with HCC after curative resection are included. The correlations between serum AFP level and clinicopathological parameters and patient survival are evaluated and compared when 400ng/mL and 20ng/mL are set as cut-off points. RESULTS: Serum AFP level is associated with more clinicopathological variables of HCC under the cut-off value of 400ng/mL, than that of 20ng/ mL. However, serum AFP level under the cut-off value of 20ng/mL is of significant prognostic impact for both overall and tumor-free survival, whereas that under 400ng/mL is not. CONCLUSIONS: The two cut-off values of preoperative AFP levels might be of different implications in cirrhotic patients with HCC after curative resection. Therefore, these might be adopted differentially in HCC.

LCSGJ-T classification, 6th or 5th edition TNM staging did not independently predict the long-term prognosis of HBV-related hepatocellular carcinoma after radical hepatectomy.

BACKGROUND: The 6th edition tumor-node-metastasis (TNM) staging (TNM-6) for hepatocellular carcinoma (HCC) was recommended. Besides, Liver Cancer Study Group of Japan (LCSGJ)-T classification has been recently proposed. However, these newly established staging systems should be further verified in different subgroups of HCC patients. The current study mainly aimed to validate the predictive power of these novel criteria in a cohort of patients with hepatitis B virus-related HCC after radical hepatectomy. As a control, the 5th edition TNM staging (TNM-5) was also evaluated. METHODS AND MATERIALS: Clinicopathological and follow-up data of consecutive 142 patients with HBV-related HCC undergoing radical hepatectomy were reviewed. The impact of variables on prognosis was determined by uni- and multivariate analyses. RESULTS: By univariate analysis, LCSGJ-T classification, TNM-6, and TNM-5 were almost significantly prognostic, except for TNM-5 for disease-free survival. Meanwhile, tumor size>or=5 cm, alpha-fetoprotein>400 ng/mL, high Edmondson-Steiner grade, presence of microvascular invasion, portal vein tumor thrombosis, satellite nodule, and resection margin

LAPTM4B-35 is a novel prognostic factor of hepatocellular carcinoma.

BACKGROUND: LAPTM4B-35 is a 35-kDa tetra-transmembrane protein overexpressed in hepatocellular carcinoma (HCC) and promotes cell survival, proliferation, and tumorigenesis. However, the potential clinical implications of LAPTM4B-35 in HCC are still unclear. This study is aimed to investigate the correlations between LAPTM4B-35 expression and prognosis in patients with HCC. METHODS: Western blot and immunohistochemistry assays were used to determine the expression of LAPTM4B-35 in HCCs and their paired noncancerous liver tissues from 65 patients. The correlations of LAPTM4B-35 expression with clinicopathological parameters were assessed by Chi-square test. Patient survival was determined by Kaplan-Meier method and log-rank test. Cox regression was adopted for multivariate analysis of prognostic factors. RESULTS: LAPTM4B-35 overexpression occurred in 76.9% of HCC tissues, while only in 4.6% of noncancerous liver tissues. Overexpression of LAPTM4B-35 was significantly associated with TNM staging and invasive tumors. Patients with higher LAPTM4B-35 expression had significantly poorer overall survival (OS) and disease-free survival (DFS) (both P < 0.001). On multivariate analysis, elevated expression of LAPTM4B-35 was found to be an independent prognostic factor for OS and DFS (P = 0.009, 0.043, respectively). CONCLUSIONS: LAPTM4B-35 overexpression is an independent prognostic factor for OS and DFS of HCC.

Expression of LAPTM4B in gallbladder carcinoma cells: the role in invasive potential.

BACKGROUND/AIMS: It was previously established that LAPTM4B-35 highly expressed in gallbladder carcinoma and being of clinicopathological and prognostic significances. However, expression of LAPTM4B gene in gallbladder carcinoma (GBC-SD), a gallbladder carcinoma cell line, and its role in invasive potential remain unclear. METHODOLOGY: Expression of LAPTM4B in GBC-SD cells was first detected. Plasmids, pcDNA3-AE (containing complete open reading frame of LAPTM4B) and Mock (pcDNA3), were transiently transfected into GBC-SD cells. Invasive phenotypes (migration and invasion) and relative molecules were then shown by transwell assay, crossing river test and Western blot analysis. RESULTS: Immunocytochemical staining revealed that LAPTM4B-35 positively expressed in cytoplasm of GBC-SD cells. But LAPTM4B-35 expression was obviously weaker in GBC-SD cells than that in BEL-7402 cells (positive control). Besides, cells transfected with pcDNA3-AE presented shorter crossing river time, less migrated and invaded cell numbers, compared with cells transfected with the Mock plasmid and parent cells. Finally, increased expressions of active uPA, MMP-9, pro MMP-2 and active MMP-2 were also observed in cells transfected with pcDNA3-AE. CONCLUSIONS: Our data suggested that LAPTM4B expressed in GBC-SD cells at a relatively low level. Forced overexpression of LAPTM4B increased invasive potential of GBC-SD cells, through modulating molecules associated with degradation of extracellular matrix.

Risk factors of recurrence and poor survival in curatively resected hepatocellular carcinoma with microvascular invasion.

BACKGROUND: Microvascular invasion (MVI) is a significant sign of the invasive property and a strong predictor of poor prognosis in hepatocellular carcinoma (HCC), a life-threatening malignancy. However, recurrence-associated and post-surgical long-term prognosis-associated factors in HCC with MVI remain unknown. OBJECTIVES: To address the abovementioned issues, based on a Chinese patient cohort with HCC after curative hepatic resection. MATERIAL AND METHODS: The patient cohort consisted of 62 consecutive patients with HCC and MVI who underwent curative hepatic resection. The associations between clinicopathologic variables and recurrence, as well as patient overall/disease-free survival, were uniand multivariately evaluated. RESULTS: Univariate χ2 test identified hepatitis B surface antigen (HBsAg) positivity, high Edmondson-Steiner grade and male gender as risk factors of recurrence, whereas Edmondson-Steiner grade and HBsAg positivity were significant or marginally significant in the multivariate stepwise logistic regression analysis. Subsequently, univariate log-rank test showed that Edmondson-Steiner grade, HBsAg positivity and Child-Pugh grade were associated with overall and/or disease-free survival. Among them, the independent prognostic impact of Edmondson-Steiner grade and HBsAg positivity for both overall and disease-free survival were proven in the multivariate Cox regression analysis. CONCLUSIONS: Our data suggested that Edmondson-Steiner grade and HBsAg positivity might serve as useful indicators of recurrence and pessimistic prognosis in HCC with MVI.

The prognostic value and non-invasive predictors of splenomegaly in cirrhotic patients with hepatocellular carcinoma following curative resection.

BACKGROUND: The prognostic value and non-invasive predictors of splenomegaly in cirrhotic patients with hepatocellular carcinoma (HCC) after curative resection remain unknown. OBJECTIVES: To investigate the prognostic value and non-invasive predictors of splenomegaly in cirrhotic patients with HCC after curative resection. MATERIAL AND METHODS: The medical records of 78 patients with HCC and liver cirrhosis who underwent curative resection were retrospectively reviewed. The influence of spleen size, measured with clinically routine ultrasonography (USG), on overall and disease-free survival was evaluated using univariate and multivariate analyses. The efficiency of some frequently used blood-derived liver function parameters and non-invasive fibrosis markers to predict splenomegaly was also assessed. RESULTS: It was shown that tumor size >5 cm, the presence of microvascular invasion, tumor-node metastasis (TNM) stage III-IVA of the tumor, spleen size >11.45 cm, and age ≤52 years were associated with poor overall survival and/or disease-free survival in univariate analyses (all p < 0.05). In multivariate analyses, spleen size was identified as an independent predictor for both overall and disease-free survival (p < 0.001 and p = 0.012, respectively). On the other hand, platelet count, aspartate aminotransferase (AST) to platelet ratio index (APRI) and Fibrosis-4 index (FIB-4) scores were significantly different between small and large spleen groups (p = 0.026, 0.003 and 0.003, respectively), while statistical differences for albumin, alanine aminotransferase (ALT), AST, total bilirubin, AST to ALT ratio (AAR), and age-platelet index (API) were not found. Using receiver operating characteristic (ROC) curves, high powers of platelet count, APRI and FIB-4 in splenomegaly prediction were confirmed. CONCLUSIONS: Splenomegaly, which can be predicted by some non-invasive variables, serves as a strong determinant for postresectional prognoses of cirrhotic patients with HCC.

Both the 5th and 6th editions of TNM staging system fail to independently predict long-term prognosis after radical hepatectomy in hepatocellular carcinoma sized > or = 5 cm.

OBJECTIVE: To validate the predictive power of the 5th and 6th editions of TNM staging system (TNM-5, TNM-6) in a Chinese patient cohort with hepatocellular carcinoma (HCC) sized > or = 5 cm after radical hepatectomy. METHODS: Consecutive 121 patients with HCC sized > or = 5 cm undergoing radical hepatectomy between January 1995 and December 2002 were included. The impact of clinicopathological variables on prognosis was determined by univariate and multivariate analyses, after excluding 2 perioperative deaths. RESULTS: In univariate analysis, TNM-5 stage did not show prognostic significance for overall or disease-free survival, as opposed to TNM-6 stage, Edmondson-Steiner grade, portal vein tumor thrombosis (PVTT), vascular invasion, satellite nodule, Child-Pugh grade, and hepatitis B surface antigen (HBsAg) positivity. When these significant variables were entered in multivariate analysis, Edmondson-Steiner grade was the sole independent prognosticator for both overall and disease-free survival, whereas Child-Pugh grade independently influenced disease-free survival. However, TNM-6 stage lost its predictive potential in multivariate analysis. CONCLUSIONS: Neither TNM-5 nor TNM-6 staging system is revealed to be independently prognostic in patients with HCC sized > or = 5 cm after radical hepatectomy. Therefore, TNM-6 calls for more support in many subsets of HCC patients.

Expression of LAPTM4B-35: a novel marker of progression, invasiveness and poor prognosis of extrahepatic cholangiocarcinoma.

LAPTM4B was proven to overexpress in hepatocellular carcinoma and relate to differentiation. We immunohistochemically investigated the expression and potential clinicopathological and prognostic significance of LAPTM4B encoded protein, LAPTM4B-35, in extrahepatic cholangiocarcinoma (EHCC) for specimens from consecutive 81 patients. LAPTM4B-35 staining was positive in cancer tissues from 59 patients (72.8%), including 12 with score 1, 22 with score 2 and 25 with score 3. No positive staining was found in non-cancer epithelia. The staining score in cancer tissues was not only significantly associated with TNM staging, histological grade, perineural and lymph node invasion (P<0.05), but also of comprehensive prognostic implications, including integrated estimation with CA19-9. These data established that LAPTM4B-35 positively expressed in a great portion of EHCC and might be a novel molecular maker of progression, invasiveness and poor prognosis.

Carbohydrate Antigen 19-9 Increases the Predictive Efficiency of α-Fetoprotein for Prognosis of Resected Hepatocellular Carcinoma.

Serum α-fetoprotein (AFP) is a classical biomarker for both diagnosis and prognosis of hepatocellular carcinoma (HCC). However, its predictive efficiency for prognosis remains unsatisfactory. This study explores whether integrating AFP and carbohydrate antigen (CA) 19-9/carcinoembryonic antigen (CEA) increase its prognostic efficiency in HCC. A total of 67 HCC patients with complete record of AFP, CA19-9, and CEA, who underwent radical hepatectomy, were included. The sole and combined evaluations for prognostic significance of the three markers were performed. In the first, it was found by one-factor analysis that AFP was a univariate prognostic indicator for disease-free survival, but not overall survival, whereas CEA and CA19-9 were not statistically significant, although the latter was of marginally predictive significance for disease-free survival. Subsequently, it was revealed that combined evaluation of AFP and CA19-9, rather than AFP and CEA, distinguished overall and disease-free survival more effectively, compared with single ones. However, this combination was not significant in multivariate Cox regression analysis, thus needing further validation, especially in large-scale prospective investigations. The addition of vascular invasion to AFP/CA19-9 combination might provide enhanced predictive power for disease-free survival. Collectively, these results preliminarily suggest that CA19-9 increases the predictive efficiency of AFP for prognosis of HCC after resection.

Overexpression of LAPTM4B-35 closely correlated with clinicopathological features and post-resectional survival of gallbladder carcinoma.

Gallbladder carcinoma (GBC) is a malignancy with dismal prognosis and unclear gene expression profile. We aimed to first present the expression of LAPTM4B-35, one product of a cancer associated gene recently cloned in hepatocellular carcinoma (HCC), and its correlation with clinicopathological features and prognosis of GBC. Immunohistochemical detection of LAPTM4B-35 was performed on samples from 75 patients with GBC. LAPTM4B-35 protein was overexpressed in 57 patients (76%) with GBC. The staining scores were significantly related to histology type, lymph node involvement, distant metastasis, Nevin staging and differentiation of GBC (P<0.05). Univariate analysis revealed that overall or disease-free survival of patients was inversely associated with its staining scores (P<0.001). Multivariate analysis showed that LAPTM4B-35 staining score was an independent prognostic marker for both overall and disease-free post-resectional survival of GBC (P=0.004 and 0.027, respectively). LAPTM4B-35 overexpressed in a majority of GBCs and correlated with clinicopathological features and post-resectional survival.

Edmondson-Steiner grade: A crucial predictor of recurrence and survival in hepatocellular carcinoma without microvascular invasio.

BACKGROUND: Microvascular invasion (MVI), an important pathologic parameter, has been proven to be a powerful predictor of long-term prognosis in hepatocellular carcinoma (HCC). However, prognostic factors in HCC without MVI remain unknown. The present study aimed to identify the risk factors of recurrence and poor post-resectional survival in this type of HCC. METHODS AND METHODS: A total of 109 patients with MVI-absent HCC underwent radical hepatectomy were enrolled. The influence of clinicopathologic variables on recurrence and patient survival was assessed using univariate and multivariate analyses. RESULTS: Chi-square test found that Edmondson-Steiner grade and satellite nodule were significantly associated with recurrence, while the former was the single marker for early recurrence. Stepwise logistic regression analysis demonstrated the independent predictive role of Edmondson-Steiner grade for recurrence. On the other hand, Edmondson-Steiner grade, serum AFP level and satellite nodule were significant for overall and disease-free survival in univariate analysis, whereas tumor size was linked to disease-free survival. Of the variables, Edmondson-Steiner grade, serum AFP level and satellite nodule were independent indicators. CONCLUSIONS: Edmondson-Steiner grade, a histological classification, carries robust prognostic implications for all the endpoints for prognosis, thus being potential to be a crucial prognosticator in HCC without MVI.

[Comparison of different clinical staging systems for hepatocellular carcinoma].

OBJECTIVE: To compare the prognostic efficacy of three clinical staging systems: the Chinese system (CS), CLIP scores, and TNM for hepatocellular carcinoma (HCC). METHODS: 107 cases diagnosed as HCC between January 2000 and October 2003 were retrospectively analyzed. Their clinical stages were decided on the basis of data in CLIP or Chinese staging system when the HCC diagnosis had been confirmed. Fifty-three of the 107 patients underwent surgical resection. The TNM stage was therefore determined according to surgico-pathological results. The survival of those patients with respective to different stages were analyzed and compared using Kaplan-Meier curves and Log rank method. RESULTS: All the patients were followed up after treatment. The 3-year survival rate in the CS Ia, Ib and CLIP 0 group were 100%. The 1- and 2-year survival rates of CS IIIa group was 68% and 51%, respectively. The 1-year survival in CS IIIb group was only 20.9%. The 1-, 2-, and 3-year survival rate in patients with a CLIP score beyond 4 was 57.1%, 49.9%, and 33.5%, respectively. In patients treated with surgical resection, the CS Ia group and TNM I group had a 3-year survival rate of 100%. The 1- and 3-year survival rate in CS IIb were 83.3% and 75.0%, in TNM III group 93.7% and 77.5%. CONCLUSION: The CLIP system may provide accurate prediction of prognosis based on early clinical features. The CS system is comparably efficient in predicting the prognosis of HCC patients, particularly in patients with severe cirrhosis.

[Results of multimodality therapy for unresectable primary liver cancer].

OBJECTIVE: To investigate the therapeutic measures for unresectable primary liver cancer (PLC). METHODS: The date of 312 unresectable primary liver cancer patients treated from January 1991 to March 2003 were retrospectively analyzed. RESULTS: Of these 312 patients, 73 were treated by cryosurgery-based combined modality therapy, 239 were treated by a TACE-oriented combined modality therapy. 289 patients except 23 were followed for a period of 2 to 156 months. The overall 1-,3- and 5-year survival rate in this series was 74. 0% , 34. 0% and 16. 7% , respectively. The 1-,3-and 5-year survival rate in the cryosurgery group was 64. 4% , 38. 4% and 27. 4% , respectively. The 1-, 3- and 5-year survival rate in the TACE group was 75. 1% , 29. 0% and 10. 0%, respectively. CONCLUSION: Treatment for the unresectable primary liver cancer should be individualized and combined with suitable therapeutic modalities.

Molecular cloning and characterization of LAPTM4B, a novel gene upregulated in hepatocellular carcinoma.

Lysosomal-associated protein transmembrane-4 beta (LAPTM4B), a novel gene upregulated in hepatocellular carcinoma (HCC), was cloned using fluorescence differential display, RACE, and RT-PCR. It contains seven exons and encodes a 35-kDa protein with four putative transmembrane regions. Both the N- and C-termini of the protein are proline-rich, and may serve as potential ligands for the SH3 domain. Immunohistochemical analysis localized the protein predominantly to intracellular membranes. Northern blot showed that the LAPTM4B mRNAs were remarkably upregulated in HCC (87.3%) and correlated inversely with differentiation status. LAPTM4B was also overexpressed in many HCC-derived cell lines. It was also highly expressed in fetal livers and certain adult normal tissues including the heart, skeletal muscle, testis, and ovary. Promoter function assays showed a distinct difference in the gene's activities between BEL7402 and HLE cell lines, suggesting that the transcription factors responsible for regulation of the gene in the two cell lines are different, and that possible negative regulatory cis-elements may exist upstream of the promoter region. It was demonstrated that the N-terminus of LAPTM4B was essential for survival of the cells. Cells harboring the full-length LAPTM4B cDNA expression clone displayed a slightly increased efficiency in colony formation. These results suggest that LAPTM4B is a potential protooncogene, whose overexpression is involved in carcinogenesis and progression of HCC. In normal cells, it may also play important roles such as regulation of cell proliferation and survival.

Expression of lysosome-associated protein transmembrane 4B-35 in cancer and its correlation with the differentiation status of hepatocellular carcinoma.

AIM: To produce high-quality polyclonal antibody to lysosome-associated protein transmembrane 4B-35 and to identify LAPTM4B-35 expression in cancer tissues and its correlation with differentiation status of hepatocellular carcinoma (HCC). METHODS: The 297 bp 5' end of LAPTM4B cDNA was obtained by PCR and inserted into prokaryotic expression vector pGEX-KG. Then the recombinant pGEX-KG-N(1-99) was transformed into E.coli JM109 to express GST-fusion protein. The fusion protein was purified by glutathione sepharose(TM) 4B agarose. The purified GST-LAPTM4B-N(1-99) was characterized by SDS-PAGE, and used to immunize rabbits. The titer and specificity of antisera were detected by ELISA and Western blot, respectively. The correlation between the expression levels of LAPTM4B-35 and the differentiation status of HCC was analyzed via Western blot. The expression of LAPTM4B-35 in HCC and other six cancer tissues was investigated via tissue chip and immunohistochemical analysis. RESULTS: About 6.2 mg of pure GST-LAPTM4B-N(1-99) was isolated from 1 L of bacteria. The GST-LAPTM4B-N(1-99) produced high titer antisera in rabbits and showed good immunity. Western blot showed specific reactions for the antibody to the LAPTM4B-35 in the total proteins from HCC tissues and BEL-7402 cells, also to the fusion protein purified or in the transformed bacteria. LAPTM4B-35 was remarkably expressed in several cancers, such as HCC, breast cancer, gastric carcinoma, lung cancer, and colon carcinoma, but not commonly expressed in esophageal cancer and rectum carcinoma. Notably, the expression levels of LAPTM4B-35 were significantly and inversely correlated to the differentiation of HCCs in a 20 case analysis. CONCLUSION: Specific polyclonal antibody (LAPTM4B-N(1-99)-pAb) to LAPTM4B-35 was produced. It identified the expression of LAPTM4B-35 in some cancer tissues originated from single layer cuboidal and columnar epithelial cells and firmly demonstrated that the expression of LAPTM4B-35 in HCC was inversely correlated with the differentiation of HCC.

Clinicopathological predictors of poor survival and recurrence after curative resection in hepatocellular carcinoma without portal vein tumor thrombosis.

Many factors associated with long-term outcome in hepatocellular carcinoma (HCC) with portal vein tumor thrombosis (PVTT) were previously identified. However, those in HCC without PVTT have not been elucidated. This study was designed to define the risk factors of poor post-surgical survival and recurrence in this subgroup of HCC. Medical records and follow-up data of consecutive 152 patients with PVTT-absent HCC underwent curative resection were reviewed. The impacts of clinical and pathological variables on patient survival and recurrence were evaluated by univariate and multivariate analyses. It was shown that Edmondson-Steiner grade, TNM stage, microvascular invasion (MVI), satellite nodule, serum AFP level, tumor size and number were significant for tumor-specific and/or tumor-free survival in univariate analysis. Among them, Edmondson-Steiner grade and TNM stage were of independent significances for both, whereas satellite nodule independently predicted tumor-free survival. In Chi-square test, Edmondson-Steiner grade, TNM stage and MVI were significantly related to overall as well as early recurrence. Stepwise logistic regression identified Edmondson-Steiner grade as the single independent predictor of both. To be summarized, variables that are associated with poor prognosis and recurrence in HCC without PVTT are all tumor-related ones. Of these, differentiation degree might be of particular importance.