Masaya Mori

The glypican 3 oncofetal protein is a promising diagnostic marker for hepatocellular carcinoma

Expression profiling of hepatocellular carcinoma has demonstrated that glypican 3 (GPC3), a heparan sulfate proteoglycan anchored to the membrane, is expressed at a markedly elevated level in hepatocellular carcinoma. In this paper, two monoclonal antibodies against GPC3, GPC3-C02 and A1836A, were confirmed to specifically recognize GPC3 molecule in cells from hepatocellular carcinoma and hepatoblastoma cell lines by immunoblotting, and both were confirmed to recognize different epitopes of the GPC3 molecule by epitope mapping. Then, we evaluated the feasibility of GPC3-immunohistochemistry in the pathological diagnosis of benign and malignant hepatocellular lesions by applying these monoclonal antibodies to formalin-fixed and paraffin-embedded specimens. The immunoreactivity turned out to be identical in the two monoclonal antibodies and was thus confirmed to represent the actual expression of the GPC3 molecule. The expression was observed in the fetal liver, but not in normal adult liver, liver cirrhosis or hepatitis except for a tiny focus of a regenerative nodule of fulminant hepatitis. Diffusely positive staining of GPC3 was observed in malignant hepatocytes in hepatoblastomas and in hepatocellular carcinomas (47/56, 84%). GPC3 expression was independent of the differentiation and size of the hepatocellular carcinoma. On the other hand, there was only weak and focal staining in low-grade (2/8) and high-grade dysplastic nodules (6/8). GPC3 immunoreactivity was detected in only one of 23 metastatic lesions of colorectal carcinoma, and its expression was entirely absent in the liver cell adenoma (0/7), carcinoid tumor (0/1), and cholangiocellular carcinoma (0/16). When compared with immunohistochemistry of hepatocyte antigen and alpha-fetoprotein, GPC3-immunohistochemistry was siginificantly much more specific and sensitive for hepatocellular carcinomas. Thus, GPC3 was confirmed to be one of the oncofetal proteins now attracting attention for their promise both as markers of hepatocellular carcinoma in routine histological examination and as targets in monoclonal antibody-based hepatocellular carcinoma therapy.

Overexpression of p53 as a possible prognostic factor in human thyroid carcinoma.

A total of 110 cases of thyroid carcinomas were examined immunohistochemically to evaluate the overexpression of mutant forms of p53 protein in the light of their relationship with their histological subtypes. A polyclonal antibody, CM-1, was applied to the routine formalin-fixed, paraffin-embedded tissues for this survey. Overall, immunohistochemically detected p53 expression confined to the nucleus was identified in 22.7% of the thyroid carcinomas. A significant difference in the positivity of p53 among histological subtypes was noted; the positivity was 11.1% of the cases in well-differentiated papillary carcinoma, 14.3% in well-differentiated follicular carcinoma, 40.9% in poorly differentiated carcinoma, and 63.6% in undifferentiated carcinoma. No immunohistochemical positivity was found in adjacent non-neoplastic tissues or in benign lesions, including follicular adenoma, adenomatous goiter, and chronic thyroiditis. These results suggest that overexpression of p53 is not a responsible factor for the oncogenesis itself, but rather that it plays a crucial role in aggressive subtypes of thyroid carcinomas. Additionally, the distinct entity of poorly differentiated carcinoma, previously categorized in the well-differentiated carcinoma under the name of papillary or follicular carcinoma, was statistically confirmed.

Stepwise participation of p53 gene mutation during dedifferentiation of human thyroid carcinomas.

The spectrum of p53 gene mutations was investigated in thyroid carcinomas with respect to histopathological classification. In all histological subtypes of thyroid carcinoma that had previously revealed positivity in immunohistochemical staining for p53 protein, single-stranded conformation polymorphism analysis and direct sequencing were performed to detect point mutations between exons 5 and 8. In well differentiated papillary and follicular carcinomas, in which we had already known that 11.1 and 14.3% of the cases, respectively, revealed p53 overexpression as determined by immunohistochemistry, genetic aberrations were undetectable. In poorly differentiated carcinoma, in which 40.9% had revealed overexpression, two of six cases revealed point mutations at codon 244 in exon 7 and at codon 278 in exon 8. In undifferentiated carcinoma, in which 63.6% had revealed overexpression, four of six cases examined showed point mutations at codon 157 in exon 5, at codon 248 in exon 7, and at codon 273 and a two-base insertion between codons 266 and 267 in exon 8. These results strongly suggest the crucial role of p53 gene aberration and protein overexpression in a biologically aggressive subtype, possibly as a stepwise participation in the process of tumor dedifferentiation in human thyroid carcinomas.

Serous cystadenoma of the pancreas with invasive growth: benign or malignant?

We describe a case of serous cystadenoma, that invaded a lymph node and adipose tissue. Preoperatively, the cystic lesion of the pancreas was diagnosed as a serous cystadenoma and subsequently the patient, a 71-yr-old woman, underwent distal pancreatectomy with splenectomy. Macroscopically, a greyish white, externally lobulated and partly ovoid tumor, measuring 12 × 8.5 × 5 cm, occupied the pancreatic body and tail extensively. In cross-section, multiple nodules were observed, which measured from 0.5 to 3 cm in diameter, were separated by hyalinized fibrous septa and were filled with numerous microcysts. Light microscopic findings were consistent with those for serous cystadenoma. At the splenic hilus, the tumor was found to have invaded the lymph node and adipose tissue. Based on the clinicopathological features of the six reported cases, including the present case (which behaved in a malignant fashion in terms of pathological findings of invasion or metastasis), serous cystadenoma should be regarded as having the potential for malignant growth.

A comparative study of postoperative complications after hepatectomy in patients with and without chronic liver disease

BACKGROUND Although hepatic resection is the most reliable treatment for hepatocellular carcinoma, impaired liver function because of cirrhosis or chronic hepatitis contributes to relatively high rates of postoperative complications. We have reviewed a series of hepatectomies at our institution and investigated risk factors for complications after hepatectomy in patients with impaired liver compared with patients with normal liver. METHODS From October 1994 to March 1998, 277 hepatectomies for hepatocellular carcinoma, cholangiocellular carcinoma, metastatic liver tumors, and other hepatic diseases were performed. In an attempt to clarify the safety of hepatectomy for the impaired liver at our institution, we did a comparative study of postoperative complications after hepatectomy in 2 groups: patients with impaired livers (187 hepatectomies) and patients with normal livers (90 hepatectomies). RESULTS Of the 277 hepatectomies, bile leakage occurred in 25 patients (16 in impaired livers vs 9 in normal livers), abdominal infection in 45 patients (30 vs 15 patients), wound infection in 13 patients (9 vs 4 patients), pleural effusion in 52 patients (35 vs 17 patients), atelectasis in 26 patients (17 vs 9 patients), pneumonia in 4 patients (3 vs 1 patients), ileus in 6 patients (3 vs 3 patients), intra-abdominal hemorrhage in 3 patients (0 vs 3 patients), and hyperbilirubinemia in 5 patients (4 vs 1 patients). Hepatic insufficiency and hospital death were not experienced in this series. The mean postoperative hospital stay was 22.9 days (23.5 vs 23.1 days), and except for intra-abdominal hemorrhage there was no statistically significant difference between the 2 groups. CONCLUSIONS Hepatectomy for the impaired liver is now as safe a procedure as for the normal liver, provided the overall guidelines outlined in our algorithm are followed.

Objective evaluation of Liver consistency to estimate hepatic fibrosis and functional reserve for hepatectomy

BACKGROUND The empiric evaluation of liver consistency is currently used to plan the surgical strategy. The aim of this study was to verify the feasibility of the objective measurement of liver consistency and to check its correlation with liver fibrosis and liver functional reserve. STUDY DESIGN Fifty-two consecutive patients who underwent hepatic resections in our department were enrolled. The indications for liver resection were hepatocellular carcinoma in 36 patients, metastatic liver tumors in 12 patients, and other conditions in 4 patients. Liver consistency was measured with a new tactile sensor. A fibrosis index was calculated as an expression of the percentage of fibrotic tissue. Liver consistency was compared with the degree of liver fibrosis observed in histologic specimens (fibrosis index) and with liver function parameters. RESULTS Liver stiffness showed a significant positive correlation with fibrosis index (r = 0.887, p < 0.0001). Liver stiffness also showed significant positive correlation with the indocyanine green test (r = 0.631, p < 0.0001) by a univariate analysis. The indocyanine green test and platelet count were independently and significantly associated with liver stiffness by a multiple regression analysis. In five patients, the liver stiffness values measured intraoperatively differed markedly from those expected from the indocyanine green test values. In these patients, the operative procedures were finally selected based on the liver stiffness measured with the tactile sensor and good clinical outcomes were obtained. CONCLUSIONS These results show for the first time that liver stiffness can be clinically assessed quantitatively by means of the tactile sensor. The tactile sensor adequately estimates liver stiffness and this estimation is well correlated with liver fibrosis and functional reserve. Liver consistency determined objectively in this manner may be useful for optimizing surgical decision making.

β-catenin (CTNNB1) S33C mutation in ovarian microcystic stromal tumors.

Microcystic stromal tumor (MCST) is a recently described subtype of ovarian tumor characterized by prominent microcystic histologic pattern and diffuse immunoreactivity for CD10 and vimentin. However, its pathobiology, particularly its histogenesis, remains largely unclear. Here, we report 2 cases of ovarian MCST, in which we have performed extensive histologic, immunohistochemical, and genetic investigations to determine its distinct nature among ovarian neoplasms. The patients were 32 and 41 years of age. Both tumors were solid and cystic masses involving the right ovary. Microscopically, tumor cells with generally bland, round-to-ovoid nuclei grew in microcystic, macrocystic, and solid patterns. Intervening thick fibrous stroma was observed. Immunohistochemically, tumor cells were diffusely and strongly positive for CD10, vimentin, and Wilms tumor 1. Furthermore, we detected aberrant nuclear expression of &bgr;-catenin protein in both cases. Of interest, mutation analyses revealed the presence of an identical point mutation, c.98C>G, in exon 3 of &bgr;-catenin (CTNNB1) in both tumors. This is an oncogenic mutation that causes replacement of serine with cysteine at codon 33, leading to the loss of a phosphorylation site in the &bgr;-catenin protein. The results of this study strongly suggest that dysregulation of the Wnt/&bgr;-catenin pathway plays a fundamental role in the pathogenesis of ovarian MCST. Finally, by comparing the immunophenotype of MCST with its histologic mimics and other ovarian sex cord-stromal tumors, we were able to identify unique features of MCST and a panel of markers useful in differential diagnosis.

Hyperactivated STAT3 in ALK-positive diffuse large B-cell lymphoma with clathrin-ALK fusion.

Anaplastic lymphoma kinase (ALK)-positive large B-cell lymphoma is a rare subtype of diffuse large B-cell lymphoma (DLBCL). Although a few cases of ALK-positive large B-cell lymphoma harbor nucleophosmin-ALK chromosomal translocation similar to ALK-positive anaplastic large cell lymphoma, most reported cases are characterized by t(2;17)(p23;q23) involving the clathrin gene. Here, we report 2 cases of ALK-positive DLBCL. The 2 cases presented similar morphologic features and immunohistochemical characteristics, that is, positivity for ALK, IgA, CD138, and MUM1; weak positivity for CD30 and CD79a; and negativity for CD20. The clathrin-ALK transcript was identified by reverse transcription-polymerase chain reaction, and the sequence was determined by direct sequencing. Recently, the essential role of STAT3 activation as well as STAT 5 activation in nucleophosmin-ALK fusion protein-mediated lymphomagenesis was reported. However, differential effects of ALK-fusion variant proteins on proliferation, transformation, and invasion properties were reported. Thus, we evaluated the phosphorylation status of STAT 3 and STAT 5, and found highly hyperphosphorylated STAT 3 on tyrosine 705 but not STAT 5 in our 2 cases of ALK-positive DLBCL with clathrin-ALK fusion. Furthermore, STAT 5A expression was not detected in either of the ALK-positive DLBCL cases, although 11 of the 36 ALK-negative DLBCL cases revealed STAT 5A expression. Expression of the antiapoptotic proteins survivin and BCL-X(L), which were believed to be the targets of STAT 3, was investigated. However, there were no significant associations between expression of survivin or BCL-X(L) and ALK positivity among the diffuse large B-cell lymphomas. In summary, similar signaling transduction mechanism involving STAT proteins seems to underlie DLBCL harboring the clathrin-ALK or nucleophosmin-ALK fusion gene.

Mucinous adenocarcinoma of the thymus: A distinct variant of thymic carcinoma

BACKGROUND Primary thymic mucinous adenocarcinoma is a recently described subtype of thymic carcinoma, which behaves aggressively. METHODS The authors analyzed the clinical and pathological findings of three cases of thymic mucinous adenocarcinoma, and reviewed five cases previously reported in the English literature. RESULTS The patients were two males and one female between the ages of 38 and 55 years. Macroscopically, the tumors were mostly solid and white to yellowish-white. Areas with a gelatinous appearance were present. Histologically, all of the tumors were adenocarcinomas with abundant mucin production, which resembled the mucinous adenocarcinomas of other organs. Malignant tumor cells in nests, tubules and cribriform structures floated in pools of extracellular mucin. In one case, associated thymic cysts were found at the periphery of the tumor. The cyst wall was partially lined by malignant mucinous epithelium, which showed transition from benign thymic epithelium. Immunohistochemically, all of the tumors showed positive immunoreactivity for cytokeratin (CK) 20 and carcinoembryonic antigen (CEA). CD5 was diffusely positive in one case, and focally positive in the other two cases. The prognoses of these cases were extremely poor, and two of the patients died within 24 months. CONCLUSION Growing evidence suggests that mucinous adenocarcinoma is a distinct morphological variant of primary thymic carcinoma. We believe that clinicians and surgical pathologists should include thymic mucinous adenocarcinoma in the differential diagnosis of mediastinal adenocarcinoma.

High-grade lung adenocarcinoma with fetal lung-like morphology: clinicopathologic, immunohistochemical, and molecular analyses of 17 cases.

Low-grade lung adenocarcinoma of fetal lung type, which is well characterized by its unique clinicopathologic and molecular features, is recognized as a distinct variant of lung cancer. In contrast, high-grade lung adenocarcinoma with fetal lung–like morphology (HG-LAFM) has not been studied widely. To characterize this subset better, we analyzed 17 high-grade adenocarcinomas with at least focal component resembling a developing epithelium in the pseudoglandular phase of the fetal lung. These rare (ca. 0.4%) carcinomas occurred predominantly in elderly men with a heavy smoking history, who showed elevated serum &agr;-fetoprotein in 4 of 5 cases tested. Histologic examination revealed a fetal lung–like component as a focal finding accounting for 5% to 60% of the total tumor volume. It was invariably admixed with tissues having a morphology not resembling that of a fetal lung. A coexisting non–fetal lung–like element was quite heterogenous in appearance, showing various growth patterns. However, clear-cell (88%), hepatoid (29%), and large cell neuroendocrine carcinoma (24%) histology seemed overrepresented. HG-LAFM was characterized immunohistochemically by frequent expression of &agr;-fetoprotein (41%), glypican-3 (88%), SALL-4 (59%), neuroendocrine markers (82%), CDX-2 (35%), and p53 (65%). HG-LAFM was molecularly heterogenous in that EGFR or KRAS mutation was observed in 22% of cases tested for both. Our data indicate that HG-LAFMs might form a coherent subgroup of lung adenocarcinomas. However, the uniformly focal nature of the fetal lung–like element, widely diverse coexisting non–fetal lung–like histology, and inhomogenous molecular profiles lead us to believe that HG-LAFM is best regarded as a morphologic pattern showing characteristic association with several clinicopathologic parameters rather than a specific tumor entity.