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(American Journal of Pathology. 2000;157:763-770.)
© 2000 American Society for Investigative Pathology

Regular Articles

ß-Catenin Mutations Are Associated with a Subset of Low-Stage Hepatocellular Carcinoma Negative for Hepatitis B Virus and with Favorable Prognosis

Hey-Chi Hsu^*, Yung-Ming Jeng^*, Tsui-Lien Mao, Jan-Show Chu, Po-Lin Lai^* and Shian-Yang Peng^§

From the Department of Pathology,^*
National Taiwan University Hospital; the Department of Pathology,
Ho-Hsin Cancer Center; the Department of Pathology,
Taipei Medical College; and the Department of General Education,^§
National Taipei College of Nursing, Taipei, Taiwan

	Abstract

Top Abstract Introduction Materials and Methods Results Discussion References

 
To better understand the role of ß-catenin mutation in hepatocellular carcinoma (HCC), we correlated the gene mutation with hepatitis virus B (HBV) and hepatitis virus C (HCV) status and the clinicopathological features in 366 patients with resected primary unifocal HCC. ß-Catenin mutations were also analyzed in 55 patients with multifocal HCC (68 tumors). Of the whole series, 57 (13.1%) of 434 tumors examined had ß-catenin mutations, 34 occurred at the serine/threonine residues of the GSK-3ß region of ß-catenin. Outside the GSK-3ß phosphorylation site, codons 32 and 34 were two mutational hot spots (17 tumors). The non-HBV-related HCC that was predominantly HCV related had a higher frequency of mutation (P < 0.00001) and more frequent mutations at codon 45 than HBV-related HCC. HBV-related HCC had a younger mean age (P < 0.00001), and higher male-to-female ratio (P < 0.003) and positive familial history of HCC (P < 0.014). Among 366 unifocal HCCs selected for clinicopathological analysis, ß-catenin mutations were associated with grade I (P = 0.005) and stage I and II HCC (P < 0.0001), and a better 5-year survival rate (P = 0.00003). These findings suggest mechanisms for ß-catenin mutations differ between HBV-related and non-HBV-related HCCs, and that ß-catenin mutation is a favorable prognostic factor related to low stage. ß-Catenin mutation was associated with nuclear expression of the protein (P < 0.00001), but we failed to detect point or large fragment deletion mutation in 39 HCCs with nuclear ß-catenin expression, presumably wild-type protein. HCCs expressing mutant nuclear ß-catenin had a better 5-year survival rate (P < 0.007), suggesting that mutant and wild-type nuclear ß-catenin proteins are not functionally equivalent and deserve more studies for further clarification.

	Introduction

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Hepatocellular carcinoma (HCC) is the leading fatal cancer in Taiwan and tumor invasion is a crucial prognostic histological factor of HCC.²⁰ HCC is closely associated with hepatitis B virus (HBV) and hepatitis C virus (HCV) infection, but the molecular mechanisms for the tumorigenesis of HBV- and HCV-related HCC are unclear. Although closely related to tumor invasion, mutation of tumor suppressor gene p53 is detected in only one-third of HCCs in Taiwan.^21,22 ß-Catenin is frequently mutated in HCC in humans^23,24 and rodents,^23,25 but the clinical implication of ß-catenin mutations in HCC needs to be clarified. In the present study, we demonstrate that ß-catenin mutation is associated with low-grade, low-stage, HBV-negative HCC, and has a more favorable prognosis.

	Materials and Methods

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Tissue Samples

From January 1983 to December 1997, 1,033 surgically resected primary and 188 recurrent HCCs were pathologically assessed at the National Taiwan University Hospital. The tissue samples were immediately cut into small pieces, snap-frozen in liquid nitrogen, and stored in deep freezers. Of these, 421 patients who already had DNA or RNA samples taken from resected primary HCCs were analyzed for ß-catenin mutations, including 366 cases of unifocal and 55 cases of multifocal HCC (68 tumors). To elucidate the clinical implication of ß-catenin mutation, the 366 cases of unifocal HCC, as previously defined,^21,22,26,27 were selected for clinicopathological analysis. Multifocal HCCs were excluded from this correlation because of incomplete sampling of the tumor nodules for genetic analysis and their variation in pathological features.

Histological Study and TNM Staging

For the convenience of comparative analyses, the tumor grade was simply divided into three groups: well (grade I), moderately (grade II), and poorly differentiated HCC (grades III and IV).

We and other investigators have shown that encapsulated HCC without liver and vascular invasion has better survival than invasive HCC, regardless of tumor size.^20,28,29 Hence, resected unifocal HCC was staged according to the staging system proposed by the International Union against Cancer (UICC, 1997), but with modification according to tumor invasion and the extent of vascular spread and listed as follows: stage I: encapsulated, without evidence of liver or vascular invasion; stage II: unencapsulated or encapsulated and with liver invasion, but without vascular invasion; stage IIIA: invasion of small vessels in the tumor capsule; stage IIIB: focal invasion of portal vein branches close to the tumor; stage IV: invasion of portal veins in distal liver (1 cm away from the tumor capsule), branches of major portal vein, common bile duct, or perforation of visceral peritoneum.

Immunohistochemistry

The ß-catenin was detected on formalin-fixed, paraffin-embedded sections by the labeled streptavidin-biotin method after antigen retrieval, as previously described,³⁰ using a monoclonal antibody against human ß-catenin (Transduction Laboratories, Lexington, KY). The ß-catenin immunostaining was scored according to the subcellular localization (along the membrane or in nucleus).

Mutational Analysis

DNA and RNA were extracted from fresh frozen liver tissues as previously described.^21,22 Primers for polymerase chain reaction were designed to amplify a 232-bp fragment of exon 3 of the ß-catenin gene,³¹ encompassing the sequence for GSK-3ß phosphorylation sites that contain activating mutations.^5,6,9,13 The primers used included sense primer: 5'-AGCTGATTTGATGGAGTTGG-3'and antisense primer: 5'-ACCAGCTACTTGTTCTTGAG-3'. DNA sequencing was performed by ABI 373 Automated Sequencer, using the ABI Prism Dye Terminator Cycle Sequencing Ready Reaction Kit (Perkin Elmer, Branchburg, NJ). Each mutation was verified in both sense and antisense directions.

To detect large fragment interstitial deletions, primers flanking exons 2 to 4 of the ß-catenin gene were used, as described by Koch et al.³² The primers for DNA templates were BCAT-3, 5'-AAAATCCAGCGTGGACAATGG-3' and BCAT-4, 5'-TGTGGCAAGTTCTGCATCATC-3', and for cDNA were BCAT-5, 5'-GGAGGAAGGTCTGAGGAGCAG-3' and BCAT-6, 5'-CGATGATGGGAAAGGTTATGC-3'. Polymerase chain reaction products of altered size were cloned to plasmid by the TA Cloning Kit (Invitrogen, Carlsbad, CA) and subjected to DNA sequencing.

To detect HBV DNA in tumor and nontumor liver samples negative for serum HBsAg, Southern blotting and/or polymerase chain reaction were carried out, as previously described.²⁶ For polymerase chain reaction, we amplified a 257-bp fragment of major S and a 433-bp fragment of C genes, respectively. The primers used were: HBV-S primer 1: 5'-ACATCAGGATTCCTAGGACCCCT-3' and HBV-S primer 2: 5'-CATAGCAGCAGGATGAAGAGGAA-3' (HBV DNA nucleotides 169 to 425), HBV-C primer 1: 5'-GCTTTGGGGCATGGACATTGACCC-3', and HBV-C primer 2: TGATAAGATAGGGGCATTTGGTGG-3' (HBV DNA nucleotides 1893 to 2325).

Follow-Up Observation

Of 366 patients who had unifocal primary HCC, 294 (80%) had been followed for more than 8 years or until death, up to 16 years. To minimize the influence of second and third recurrent primary HCC,^21,22,26 the endpoint for follow-up was set at 5 years.

Statistical Analyses

The analyses were carried out using the Statistica for Window software (Statsoft, Inc., Chicago, IL). We used two-tailed chi-square and Fisher exact tests for univariate analysis. The cumulative survival after tumor removal was calculated with log-rank test. P values <0.05 were considered statistically significant.

	Results

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Clinical Features of 421 Study Cases with Primary HCC

There were 339 male and 82 female patients from 10 to 88 years of age. HBsAg was detected in the sera in 280 patients (66.5%; 69.6% in the whole series of 1033 cases), and HBV genome in liver tissues in 29 HBsAg-seronegative cases. For the convenience of comparison, these 309 patients, including 44 cases also positive for anti-HCV, or co-infection, were regarded to have HBV-related HCC. In the remaining 112 non-HBV-related HCC patients, 95 (85%) were positive for anti-HCV and 17 were negative for HBV and HCV. Of these 17 cases, none was alcoholic, four had liver cirrhosis, four had chronic persistent hepatitis, and nine had normal liver histology. HBV-related HCC had younger mean age (52.7 ± 13.6 years versus 63.9 ± 9.4 years, P < 0.00001), higher male-to-female ratio (84.1% versus 70.5%, P < 0.003), more frequent serum alpha-fetoprotein (AFP) elevation above 320 ng/ml (49.5% versus 32.1%, P = 0.002), and positive familial history of HCC (14.9% versus 5.4%, P < 0.014). Liver cirrhosis was found in 40.5% and 43.8% of HBV-related and non-HBV-related HCC, respectively.

Frequency and Mutational Hotspots of ß-Catenin Mutation

To analyze the pattern and frequency of ß-catenin mutations, both unifocal and multifocal HCCs were included. ß-Catenin mutations were detected in 57 (13.1%) out of 434 tumors taken from 421 HCC patients, 30 mutations in HBV-related and 27 in non-HBV-related HCC (Figures 1 and 2) . Thirty-four mutations (59.6%) were at the phosphorylation sites for the GSK-3ß binding, including five at codon 33, five at codon 37, nine at codon 41, and 15 at codon 45. Nineteen mutations (33.3%) occurred outside the phosphorylation sites, including 10 at codon 32, seven at codon 34, and two novel mutations at codon 36. Deletion mutations were found in four tumors.

View larger version (72K): [in this window] [in a new window]   Figure 1. Mutations of the ß-catenin gene in hepatocellular carcinoma. Representative mutations leading to amino acid substitutions are shown. A–F: Point mutations at codons 32, 34, 41, and 45: Asp32 Tyr (A), Asp32 Gly (B), Gly34 Val (C), Gly34 Glu (D), Thr41 Ala (E), and Ser45 Pro (F). A small deletion of codons 45–48 (G) and a large segment deletion of codons 10–141 (H) are indicated.

View larger version (12K): [in this window] [in a new window]   Figure 2. Illustration of the location of 57 mutations of the ß-catenin gene in hepatitis B virus-positive HBV(+) and hepatitis B virus-negative HBV(-), primary unifocal and multifocal hepatocellular carcinoma. Of the HBV(-) HCC, 21 (87.5%) of 24 tumors were taken from patients positive for anti-HCV. The amino acid substitutions of 53 point mutations in exon 3 are indicted at the top, whereas four deletion mutations, designated as , are indicated at the bottom. Bold type indicates potential phosphorylation sites on threonine and serine residues. *, designates HBV(+) patients who were also positive for anti-HCV in sera. Single-letter abbreviations for the amino acid residues are: A, Ala; C, Cys; D, Asp; E, Glu; F, Phe; G, Gly; H, His; I, Ile; K, Lys; L, Leu; M, Met; N, Asn; P, Pro; Q, Gln; R, Arg; S, Ser; T, Thr; V, Val; W, Trp; and Y, Tyr.

ß-Catenin mutations occurred more frequently in non-HBV-related HCCs in both unifocal and multifocal HCCs (P < 0.00001), regardless of presence or absence of anti-HCV (Table 1) . In addition to the difference in frequency, the pattern of ß-catenin mutations also differed between HBV-related and non-HBV-related HCCs.

Of the 366 unifocal HCCs selected for clinicopathological analysis, the tumor was 2 cm in 31, 2.1 to 5 cm in 143, and larger than 5 cm in 192. Stage I, stage II, stage IIIA, stage IIIB, and stage IV HCCs had 80, 83, 67, 50, and 86 cases, respectively. There was a close correlation between tumor stage and patient’s outcome (P < 0.000001) (Figure 3) . ß-Catenin mutations occurred more frequently in older patients (P = 0.0009), but inversely correlated with serum AFP elevation (P = 0.0001) (Table 3) . Histologically, ß-catenin mutation was associated with grade I HCC (P = 0.005). Moreover, HCC with ß-catenin mutations had a significantly higher frequency of stage I and II HCCs, but less commonly high-stage tumor with portal vein invasion (stage IIIB and IV HCCs; P < 0.0001) (Table 3) . ß-Catenin mutations did not correlate with tumor size (Table 3) and liver cirrhosis (data not shown). HCC with ß-catenin mutation had a significantly better 5-year survival rate than HCC without the gene mutation (P = 0.00003) (Figure 4) .

View larger version (19K): [in this window] [in a new window]   Figure 3. Cumulative survival curves for 366 patients with unifocal hepatocellular carcinoma (HCC) according to tumor stage. The 5-year survival rates for stage I, II, IIIA, IIIB, and IV HCC are 63%, 43%, 22%, 17%, and 5%, respectively (log-rank test, P < 0.000001).

View larger version (17K): [in this window] [in a new window]   Figure 4. Cumulative survival curve for 366 patients of primary unifocal hepatocellular carcinoma (HCC). HCC with ß-catenin mutations had a significantly better 5-year survival rate (P = 0.00003).

ß-Catenin protein was examined by immunohistochemical stain in 282 cases with unifocal HCC. Of these, 212 expressed membranous ß-catenin alone, whereas nuclear accumulation of ß-catenin, ranging from diffuse to scattered cells, was detected in 70 cases (Figure 5) . ß-Catenin mutation was significantly associated with nuclear expression of the protein, 83.8% (or 31 out of 37) versus 15.9% (or 39 out of 245) (P < 0.00001). We failed to detect point or large fragment deletion mutations of ß-catenin gene in 39 tumors with nuclear expression of ß-catenin, presumably wild-type protein. HCCs with mutant nuclear ß-catenin expression had significantly higher 5-year survival rate than HCCs with wild-type nuclear ß-catenin expression (66% versus 34%, P < 0.007) (Figure 6) .

View larger version (125K): [in this window] [in a new window]   Figure 5. Expression of ß-catenin protein. A: Diffuse staining of ß-catenin along the tumor cell membranes of HCC is shown. B: Membranous staining of ß-catenin of mixed positive (right) and negative areas (left) is shown. C: Abundant tumor cell nuclei are strongly positive for ß-catenin. D: Scattered tumor cells are positive for nuclear ß-catenin (arrows). Immunohistochemical stain for ß-catenin; original magnification x340 (A–D).

View larger version (19K): [in this window] [in a new window]   Figure 6. Cumulative survival curve for primary unifocal hepatocellular carcinoma (HCC) that had nuclear ß-catenin protein expression. HCC with nuclear ß-catenin protein expression and ß-catenin mutation (n = 31) had a better 5-year survival rate than HCC with nuclear ß-catenin protein expression but without mutation of the gene (n = 39) (P < 0.007, log-rank test).

	Discussion

Top Abstract Introduction Materials and Methods Results Discussion References

Of the 57 mutations, 34 (59.6%) occurred at the serine/threonine residues in the GSK-3ß phosphorylation consensus motif of ß-catenin,³⁹ four had deletion mutations, and 19 (33.3%) had mutations outside these phosphorylation residues: at codons 32 (10 tumors), 34 (seven tumors), and 36 (two tumors), flanking the serine residues at codons 33 and 37. Our findings confirm that codons 32 and 34 are mutational hot spots in both human and animal HCC.^23,24 The frequent mutations at codons 32 and 34 that are uncommon in colorectal carcinoma,⁵ melanoma,⁹ and endometrial carcinoma¹⁴ provide a novel ß-catenin activating mechanism in human and murine HCC.^23,24 These findings indicate that different types of ß-catenin mutations reflect different etiologies of carcinogenesis in specific tissue.²⁴

Despite the frequent mutations of ß-catenin in various types of human cancer, the clinical implication is not fully understood. To elucidate the role of ß-catenin mutations, we selected the 366 cases of unifocal HCC for further statistical analysis. In contrast to the observations of Legoix et al,³⁶ we found that mutations of ß-catenin in HCC correlated with several important clinicopathological features of HCC. In addition to having an older mean age (P = 0.0009), HCC with ß-catenin mutations correlated inversely with serum AFP elevation (P < 0.00001), and positively with grade I HCC (P = 0.005). Moreover, HCC with ß-catenin mutations was mostly stage I and II HCC (52.5%), whereas HCC without ß-catenin mutations often had high-stage tumor with portal vein invasion (stage IIIB and IV HCC, 40.7%) (P < 0.0001). In this study, we demonstrated that HCC with ß-catenin mutation had a more favorable outcome (P = 0.00003). This finding was in accord with the results that tumor stage closely correlated with prognosis (P < 0.000001). The reasons for the inverse correlation of ß-catenin mutation with tumor aggression are not clear. Nevertheless, our results are in accord with the observations that mutations of ß-catenin were observed in various types of benign human neoplasms,^18,19 and low-stage endometrioid ovarian carcinoma with good prognosis.^40,41 Large fragment deletion of ß-catenin was detected in two out of five hepatocellular adenomas that is not a precursor of HCC (Hsu, unpublished data). N-terminally deleted ß-catenin is associated with differentiated morphology and reduced invasiveness and metastasis.⁴² Whether ß-catenin mutations that are predominantly point mutations in HCC also possess invasion/metastasis suppressive potential in human HCC deserves more studies for clarification.

In the nucleus, ß-catenin regulates transcription of target genes by binding the Tcf-Lef family of DNA binding proteins,^3-8 and enhanced wild-type ß-catenin expression is critical for the tumor suppressive effect of APC.⁵ We found that ß-catenin mutations were strongly associated with nuclear ß-catenin expression (P < 0.00001), confirming that mutations of ß-catenin lead to decreased degradation and accumulation of the protein in the nucleus.^3,4,11 In addition to the close association of nuclear ß-catenin expression with the gene mutation, reflecting mutant ß-catenin, we also demonstrated nuclear ß-catenin expression in 39 HCCs that had no point or large fragment deletion mutation of the gene, presumably reflecting wild-type ß-catenin. Moreover, HCC with mutant nuclear ß-catenin expression had a significantly higher 5-year survival rate (P < 0.007). These findings suggest that mutant and wild-type nuclear ß-catenin proteins are not functionally equivalent. Because ß-catenin mutation is associated with nuclear protein accumulation, our results also suggest that it is the qualitative change of ß-catenin protein, rather than its quantity change in the nucleus, that is important for the better survival of HCC patients with the gene mutation. More studies are hence warranted to better understand the molecular mechanisms of the potential tumor invasion/metastasis suppressive effect of mutant ß-catenin in HCC.

	Footnotes

 
Address reprint requests to Dr. Hey-Chi Hsu, Department of Pathology, National Taiwan University Hospital, 7 Chung-Shan South Road, Taipei, Taiwan. E-mail: heychi{at}ha.mc.ntu.edu.tw

Supported by the National Science Council of the Republic of China, Taiwan (NSC88–0419-002 to H. C. H.), and the Hauman Enterprises Co., Ltd., Taipei, Taiwan (to H. C. H.)

Accepted for publication May 25, 2000.

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				W. Cheng, C.-J. Tseng, T. T.C. Lin, I. Cheng, H.-W. Pan, H.-C. Hsu, and Y.-M. Lee Glypican-3-mediated oncogenesis involves the Insulin-like growth factor-signaling pathway Carcinogenesis, July 1, 2008; 29(7): 1319 - 1326. [Abstract] [Full Text] [PDF]

				T. Yamashita, M. Forgues, W. Wang, J. W. Kim, Q. Ye, H. Jia, A. Budhu, K. A. Zanetti, Y. Chen, L.-X. Qin, et al. EpCAM and {alpha}-Fetoprotein Expression Defines Novel Prognostic Subtypes of Hepatocellular Carcinoma Cancer Res., March 1, 2008; 68(5): 1451 - 1461. [Abstract] [Full Text] [PDF]

				P-M Chiang, R-H Yuan, H-C Hsu, T-L Mao, R-H Hu, P-L Lai, and Y-M Jeng Frequent nuclear expression of {beta}-catenin protein but rare {beta}-catenin mutation in pulmonary sclerosing haemangioma J. Clin. Pathol., March 1, 2008; 61(3): 268 - 271. [Abstract] [Full Text] [PDF]

				M.-C. Su, C.-C. Wang, C.-C. Chen, R.-H. Hu, T.-H. Wang, H.-L. Kao, Y.-M. Jeng, and R.-H. Yuan Nuclear Translocation of {beta}-Catenin Protein but Absence of {beta}-Catenin and APC Mutation in Gastrointestinal Carcinoid Tumor Ann. Surg. Oncol., December 1, 2006; 13(12): 1604 - 1609. [Abstract] [Full Text] [PDF]

				Y.-C. Hsu, H.-H. Fu, Y.-M. Jeng, P.-H. Lee, and S.-D. Yang Proline-Directed Protein Kinase FA Is a Powerful and Independent Prognostic Predictor for Progression and Patient Survival of Hepatocellular Carcinoma J. Clin. Oncol., August 10, 2006; 24(23): 3780 - 3788. [Abstract] [Full Text] [PDF]

				A. N.M. Fischer, B. Herrera, M. Mikula, V. Proell, E. Fuchs, J. Gotzmann, R. Schulte-Hermann, H. Beug, and W. Mikulits Integration of Ras subeffector signaling in TGF-{beta} mediated late stage hepatocarcinogenesis Carcinogenesis, May 1, 2005; 26(5): 931 - 942. [Abstract] [Full Text] [PDF]

				R.-H. Yuan, Y.-M. Jeng, H.-L. Chen, F.-J. Hsieh, C.-Y. Yang, P.-H. Lee, and H.-C. Hsu Opposite Roles of Human Pancreatitis-Associated Protein and REG1A Expression in Hepatocellular Carcinoma: Association of Pancreatitis-Associated Protein Expression with Low-Stage Hepatocellular Carcinoma, {beta}-Catenin Mutation, and Favorable Prognosis Clin. Cancer Res., April 1, 2005; 11(7): 2568 - 2575. [Abstract] [Full Text] [PDF]

				D. F. Calvisi and S. S. Thorgeirsson Molecular Mechanisms of Hepatocarcinogenesis in Transgenic Mouse Models of Liver Cancer Toxicol Pathol, January 1, 2005; 33(1): 181 - 184. [Abstract] [Full Text] [PDF]

				Y.-S. Jou, C.-S. Lee, Y.-H. Chang, C.-F. Hsiao, C.-F. Chen, C.-C. Chao, L. S.H. Wu, S.-H. Yeh, D.-S. Chen, and P.-J. Chen Clustering of Minimal Deleted Regions Reveals Distinct Genetic Pathways of Human Hepatocellular Carcinoma Cancer Res., May 1, 2004; 64(9): 3030 - 3036. [Abstract] [Full Text] [PDF]

				Y.-M. Jeng, S.-Y. Peng, C.-Y. Lin, and H.-C. Hsu Overexpression and Amplification of Aurora-A in Hepatocellular Carcinoma Clin. Cancer Res., March 15, 2004; 10(6): 2065 - 2071. [Abstract] [Full Text] [PDF]

				M. Fiorentino, A. Altimari, M. Ravaioli, E. Gruppioni, E. Gabusi, B. Corti, M. Vivarelli, P.-P. Bringuier, J.-Y. Scoazec, W. F. Grigioni, et al. Predictive Value of Biological Markers for Hepatocellular Carcinoma Patients Treated with Orthotopic Liver Transplantation Clin. Cancer Res., March 1, 2004; 10(5): 1789 - 1795. [Abstract] [Full Text] [PDF]

				B. Yang, M. Guo, J. G. Herman, and D. P. Clark Aberrant Promoter Methylation Profiles of Tumor Suppressor Genes in Hepatocellular Carcinoma Am. J. Pathol., September 1, 2003; 163(3): 1101 - 1107. [Abstract] [Full Text] [PDF]

				R. B. Birrer, D. Birrer, and J. V. Klavins Hepatocellular Carcinoma and Hepatitis Virus Ann. Clin. Lab. Sci., January 1, 2003; 33(1): 39 - 54. [Abstract] [Full Text] [PDF]

				S.-H. Liu, C.-Y. Lin, S.-Y. Peng, Y.-M. Jeng, H.-W. Pan, P.-L. Lai, C.-L. Liu, and H.-C. Hsu Down-Regulation of Annexin A10 in Hepatocellular Carcinoma Is Associated with Vascular Invasion, Early Recurrence, and Poor Prognosis in Synergy with p53 Mutation Am. J. Pathol., May 1, 2002; 160(5): 1831 - 1837. [Abstract] [Full Text] [PDF]

				S. C. Abraham, T.-T. Wu, R. H. Hruban, J.-H. Lee, C. J. Yeo, K. Conlon, M. Brennan, J. L. Cameron, and D. S. Klimstra Genetic and Immunohistochemical Analysis of Pancreatic Acinar Cell Carcinoma : Frequent Allelic Loss on Chromosome 11p and Alterations in the APC/{beta}-Catenin Pathway Am. J. Pathol., March 1, 2002; 160(3): 953 - 962. [Abstract] [Full Text] [PDF]

				S. Inagawa, M. Itabashi, S. Adachi, T. Kawamoto, M. Hori, J. Shimazaki, F. Yoshimi, and K. Fukao Expression and Prognostic Roles of {beta}-Catenin in Hepatocellular Carcinoma: Correlation with Tumor Progression and Postoperative Survival Clin. Cancer Res., February 1, 2002; 8(2): 450 - 456. [Abstract] [Full Text] [PDF]

				S. C. Abraham, E. A. Montgomery, F. M. Giardiello, and T.-T. Wu Frequent {beta}-Catenin Mutations in Juvenile Nasopharyngeal Angiofibromas Am. J. Pathol., March 1, 2001; 158(3): 1073 - 1078. [Abstract] [Full Text] [PDF]

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