Clinicopathologic study on clear cell hepatocellular carcinoma

In order to study the clinicopathologic characteristics of the clear cell variant of hepatocellular carcinoma (HCC), 215 consecutive cases measuring less than 5 cm in diameter were reviewed. The cases were divided into clear cell HCC (20 cases); focal clear cell HCC (77 cases); and non-clear cell HCC (118 cases). Clinical and pathological findings were compared among these groups. Clear cell HCC was moderately differentiated in 80% of cases and the incidence was not related to tumor size. The male to female ratio was 2.3:1, lower than the 6.9:1 of non-clear cell HCC. The association rate with liver cirrhosis was 90%, higher than the 59.3% of non-clear cell HCC. Three- and five-year survival rates, and no recurrence time were 54.5%, 33.3%, and 564 days, respectively, lower than the findings of 74.3%, 46.1%, and 770 days for non-clear cell HCC. But there is no significant difference in prognosis between both groups. Ultrastructurally, clear cells showed abundant glycogen storage and a variable degree of fat vacuoles, with a marked reduction of the number and size of organelles in the 8 cases examined. Non-clear cells of focal clear cell and non-clear cell HCC showed a moderate degree of glycogen storage in 85.7% and 28.6% of the seven cases examined from each group, with significant difference. It was concluded that clear cell HCC occurs mostly in the moderately differentiated form and is characterized by high female prevalence, high rate of association with liver cirrhosis, and has no significant difference in prognosis compared with non-clear cell HCC.     

Relationship between p53 overexpression and the proliferative activity in hepatocellular carcinoma

Relationship between p53 protein overexpression and clinicopathological findings and the proliferative activity was studied in 50 cases of hepatocellular carcinoma (34 biopsy and 16 surgically resected cases) using immunohistochemistry. Overexpression of p53 was observed in 26.5% of biopsy cases and 31. 3% of surgically resected cases. Investigation of the relationship between the p53-positive rate and the clinical stage of HCC showed that it was significantly higher in Stage IV (the most advanced cancer; 54.5%) than in Stage I/II/III (13.0%) (p<0.05). Examination of the relationship between the p53-positive rate and tumor differentiation in the biopsy cases showed that p53 was positive in 9.1% of well differentiated carcinomas, 21.4% of moderately differentiated carcinomas, and 55.6% of poorly differentiated carcinomas, indicating that p53 positivity increased as tumors became less differentiated. The p53-positive rate of poorly differentiated carcinoma (55.6%) was significantly higher than that of well and moderately differentiated carcinoma (16.0%) (p<0.05). In the surgically resected cases, p53 overexpression tended to be more frequent in the less differentiated parts of each tumor nodule. In cases with nodule in nodule pattern of HCC, the p53-positive rate was different among nodules with the same level of differentiation. Examination of tumor cell proliferative activity using the proliferating cell nuclear antigen L.I. showed that this indicator was significantly higher in the p53-positive tumors than in the p53-negative tumors (52.7+/-32.4% vs. 32.4+/-15.3%: p<0.05). These results suggest that p53 overexpression may be involved in determining the dedifferentiation and the proliferative activity of HCC. Examination of the surgically resected cases confirmed that p53 overexpression became heterogeneous during the multistep carcinogenesis and growth process of HCC, which is considered to develop from a single cell. This finding suggests that p53 overexpression may be involved in tumor progression.     

Hepatocellular Carcinoma Presenting Well Differentiated,Normotrabecular Patterns in Peripheral or Metastatic Loci

In a series of 103 surgically resected hepatocellular carcinomas (HCCs), the tumor boundaries were rather clearly defined in 85 cases (82.5%). Twelve tumors had an ill-defined margin because adjacent pseudolobules were replaced by HCC with a well differentiated growth pattern (group Al). This peripheral pattern was unrelated to the predominant histologic type of the major tumor nodules. Despite its close resemblance to normal hepatic parenchyma, we were able to determine the area of the normotrabecular HCC on the basis of, differences in stain-ability, variable nuclear crowding and frequent microacinar formation. Moreover, scattered occurrence of nuclear atypia and midtrabecular conversion were also observed within the normotrabecular HCC. In an additional 6 cases, a band like zone of normotrabecular HCC was evident peripherally around the primary lesion (group A2). Fourteen other cases (13.6%) had intrahepatic metastatic lesions formed either within the portal vein or in pseudolobules. Of these, 5 cases showed intrapseudo-lobular metastases with a normotrabecular pattern (group B). For precise evaluation of surgical HCC specimens, it seems to be particularly important to realize that HCCs sometimes have a highly differentiated appearance in a locus of peripheral invasion as well as intrahepatic metastasis. Acta Pathol Jpn 40: 887–893, 1990.     

Hepatocellular carcinoma presenting well differentiated, normotrabecular patterns in peripheral or metastatic loci. Analysis of 103 resected cases

In a series of 103 surgically resected hepatocellular carcinomas (HCCs), the tumor boundaries were rather clearly defined in 85 cases (82.5%). Twelve tumors had an ill-defined margin because adjacent pseudolobules were replaced by HCC with a well differentiated growth pattern (group A1). This peripheral pattern was unrelated to the predominant histologic type of the major tumor nodules. Despite its close resemblance to normal hepatic parenchyma, we were able to determine the area of the normotrabecular HCC on the basis of, differences in stain-ability, variable nuclear crowding and frequent microacinar formation. Moreover, scattered occurrence of nuclear atypia and midtrabecular conversion were also observed within the normotrabecular HCC. In an additional 6 cases, a band-like zone of normotrabecular HCC was evident peripherally around the primary lesion (group A2). Fourteen other cases (13.6%) had intrahepatic metastatic lesions formed either within the portal vein or in pseudolobules. Of these, 5 cases showed intrapseudolobular metastases with a normotrabecular pattern (group B). For precise evaluation of surgical HCC specimens, it seems to be particularly important to realize that HCCs sometimes have a highly differentiated appearance in a locus of peripheral invasion as well as intrahepatic metastasis.     

Angiogenesis in liver cirrhosis and hepatocellular carcinoma

BACKGROUND: Angiogenesis has been well documented in hepatocellular carcinoma (HCC). As liver cirrhosis is considered preneoplastic condition, the aim of this study was to evaluate the process of angiogenesis using CD 34 as an endothelial cell marker in normal liver, cirrhosis and HCC. MATERIALS AND METHODS: A total of 111 cases were included in this study, which consisted of 30 cases each of normal liver and cirrhosis that were all autopsy cases. Twenty-one cases of HCC included 10 autopsy specimens, nine surgically resected specimens and two liver biopsies. Remaining were 30 cases of metastasis to the liver, which included 20 autopsy specimens, one surgically resected specimen and nine liver biopsies. The patients were between the age range from 17 to 80 years with 70 males and 11 females. Paraffin-embedded liver sections of all these cases were stained routinely by hematoxylin-eosin stain, while immunohistochemistry for CD 34 was performed for expression of endothelial cells. The positivity of CD 34 staining was evaluated by counting in 10 high-power field, grading was done from 0 to 4 and compared between normal liver, cirrhosis and HCC and metastasis. RESULTS: CD 34 was positive in 16/30 (53.3%) cases of cirrhosis, 18/21 (85%) cases of HCC and 26 (86.6%) of metastasis to the liver. None of the normal liver showed any positivity. Grade 3 to 4 positivity was seen in 4/16 (25%) and 13/18 (72%) cases of cirrhosis and HCC, respectively. Amongst these, 10 were moderately differentiated, one well differentiated and rest two were fibrolamellar and sarcomatoid variants of HCC. CONCLUSION: Over expression of endothelial cell marker CD 34 with gradual progression was found from normal liver to cirrhosis to HCC and metastasis. Understanding of this process of angiogenesis might help in the design of efficient and safe antiangiogenic therapy for these liver disorders.     

非小细胞肺癌中Axin与β-连环素异常表达的关系

目的 探讨Axin（axis inhibition protein）和β-连环素（β-catenin）在非小细胞肺癌（NSCLC）中的表达、β-catenin突变及它们与各临床病理因素的关系。方法 采用免疫组织化学（SP法）和直接测序方法对100例NSCLC标本中Axin和β-catenin的表达和β-catenin基因突变进行了检测。结果 β-catenin的细胞膜表达降低率为80．0％,细胞核表达率为26．0％。高、中分化和低分化NSCLC中,β-catenin的表达降低率分别为70．0％（35／50）和90．0％（45／50）,差异有统计学意义（P=0．012）。有淋巴结转移和无淋巴结转移NSCLC中β-catenin表达降低率分别为87．3％（48／55）和71．1％（32／45）,差异有统计学意义（P=0．044）。Axin的阳性表达率为48．0％,高、中分化和低分化NSCLC中,Axin的阳性率分别为60．0％（30／50）和36．0％（18／50）,差异有统计学意义（P=0．016）。在β-catenin核表达阳性的病例中,Axin阳性表达率为15．4％（4／26）,低于β-catenin核表达阴性的病例（59．5％,44／74）（P〈0．001）。100例肺癌新鲜标本中未发现β-catenin基因第三外显子突变。结论 β-catenin的膜表达降低与NSCLC的低分化和淋巴结转移相关,Axin的表达与NSCLC的低分化和β-catenin的细胞核蓄积负相关。β-catenin基因第三外显子的突变可能不是NSCLC中β-catenin蛋白异常表达的主要原因。     

Diagnostic utility of CD10 in differentiating hepatocellular carcinoma from metastatic carcinoma in fine-needle aspiration biopsy (FNAB) of the liver

The differential diagnosis between hepatocellular carcinoma (HCC) and metastatic carcinoma, especially in moderate-poorly differentiated (MPD) HCC and poorly differentiated carcinoma, can be challenging in fine-needle aspiration biopsy (FNAB) of the liver. Recent studies demonstrate that canalicular staining for CD10 appears to be a highly specific marker for hepatocytic differentiation. The objective of this study was to test the utility of CD10 in differentiating HCC from metastatic carcinoma in FNAB of the liver. Formalin-fixed, paraffin-embedded cell blocks of 55 cases (22 HCC, 23 metastases, and 10 benign hepatic lesions) of FNAB of the liver were immunostained using monoclonal antibody against CD10, with microwave oven antigen retrieval, followed by a standard ABC method. Nineteen (86%) of 22 HCC cases were positive for CD10 with a canalicular staining pattern. Among them, 9 (82%) of 11 well-differentiated (WD) HCC and 10 (91%) of 11 MPD HCC were positive for CD10. Three (13%) of 23 metastatic carcinomas were positive for CD10, demonstrating a contrasting cytoplasmic and membranous staining pattern. The three positive cases were metastatic renal cell carcinoma (RCC), choriocarcinoma, and adenocarcinoma of the lung. All 10 cases of benign hepatic lesions showed positivity for CD10 with a canalicular and focal membranous staining pattern. In conclusion, CD10 appears to be a useful marker in discriminating between HCC and metastatic carcinoma when applied to FNAB of the liver. CD10 does not provide discrimination between WD HCC and benign hepatocytes.     

Morphological clues for the diagnosis of small hepatocellular carcinomas

Summary Histological features of 44 cases of small hepatocellular carcinoma (HCC) were examined and compared with those of large regenerative nodules. The highly differentiated type of HCC most often occurred in nodules which were less than 2 cm in diameter. Noticeably, in 9 out of 15 such cases (60.0%), tumour cells were arranged in trabeculae of almost normal thickness (normotrabecular pattern). These trabeculae, however, showed variable nuclear crowding, occasional microacinar formation, and increase in cytoplasmic basophilia. It is emphasized that the presence of this triad may be a very reliable indicator for the histological identification of early HCC, especially in examining limited material such as a biopsy specimen. However, cellular and structural atypia becomes more prominent in nodules which are larger than 2 cm.     

Fine-needle aspiration cytology of Hürthle cell carcinoma of the thyroid

Specific criteria for the diagnosis of fine-needle aspiration (FNA) of Hürthle Cell Carcinoma (HCC) have rarely been discussed in the literature. A retrospective review of 35 FNA cases with the diagnosis of Hürthle cell lesion or Hürthle cell neoplasm was performed. In each case, there was a subsequent surgical excision. The FNA specimens were divided according to histologic diagnoses as HCC (12 cases), Hürthle cell adenoma (HCA) (14 cases), and benign nonneoplastic Hürthle cell lesions (BNHCL) (9 cases). Each case was examined using a semiquantitative scoring system for the following 11 features: presence or absence of colloid, lymphocytes, and transgressed blood vessels (each scored 0 or 1); the percentage of nuclear enlargement, small cell dysplasia, large cell dysplasia, nuclear crowding, and cellular dyshesion (each scored 0-3); and age, gender, and size of lesion. When diagnosed by FNA as either Hürthle cell neoplasm or Hürthle cell lesion, males were much more likely to have malignant tumors than females. Statistically significant cytologic features that favored malignant (HCC) over benign lesions (HCA and BNHCL) included small cell dysplasia, large cell dysplasia, nuclear crowding, and cellular dyshesion. The presence of colloid and lymphocytes favored a benign lesion. Nuclear enlargement and large tumor size are significantly more common in neoplasms than BNHCL.     

Peritoneal seeding of hepatocellular carcinoma: clinicopathological characteristics of 17 autopsy cases

Peritoneal seeding (PS) of hepatocellular carcinoma (HCC) is uncommon, and the clinicopathological features are poorly understood. A total of 181 autopsy cases of HCC, including 171 cases with detailed clinical information, was investigated for PS and evaluated. PS was identified in 17 cases (9.4%), and was locally (70.6%) or entirely (29.4%) distributed in the peritoneal cavity, involving the diaphragm (76.5%), omentum (47.1%), or alimentary tract serosa (47.1%). Compared with primary HCC, PS showed similar or slightly undifferentiated features (88.2%) and exhibited more differentiated features (11.8%). In 15 cases (88.2%) of HCC with PS, primary HCC showed membranous β-catenin immunoreactivity. However, in five cases (33.3%), respective PS lost this immunoreactivity. PS was significantly associated with rupture of HCC (P= 0.012), direct diaphragmatic invasion (P= 0.001), and lymph node metastasis (P < 0.001), indicating these are high risk factors for PS; there was no significant association with a past history of percutaneous fine-needle biopsy, percutaneous ethanol injection and/or radiofrequency ablation (P= 0.97), or metastasis to lung (P= 0.13), bone (P= 0.71), or adrenal gland (P= 0.79). PS can infrequently proliferate aggressively with more differentiated features. Loss of membranous β-catenin expression may be associated with PS of HCC.     

Histological study of PIVKA-II expression in hepatocellular carcinoma and adenomatous hyperplasia

Although serum concentration of protein induced vitamin K absence or antagonist II (PIVKA-II) has been widely used for diagnosing hepatocellular carcinoma (HCC), little information is available concerning tissue PIVKA-II as an immunohistochemical marker for liver histology. In this study, we examined the expression of PIVKA-II in precancerous nodules (adenomatous hyperplasia) and various differentiation grades of HCC by immunohistochemical study using the monoclonal anti-PIVKA-II antibody (MU-3). We examined the relationship between tissue PIVKA-II staining and serum PIVKA-II level, tumor histology and tumor size. PIVKA-II was mainly detected in the cytoplasm of the HCC cells. The positive rates of PIVKA-II were as follows: adenomatous hyperplasia (AH), 0% (0/9); well-differentiated HCC, 65% (15/23); moderately differentiated HCC, 85% (22/26); poorly differentiated HCC, 54% (7/13). The expression of tissue PIVKA-II staining in moderately differentiated HCC was significantly higher than in well- or poorly differentiated HCC, whereas the serum PIVKA-II level in poorly differentiated HCC was higher than well- or moderately differentiated HCC. There was no relationship between the expression of PIVKA-II in cancer tissues and serum levels of PIVKA-II. Immunohistochemical studies revealed that PIVKA-II was expressed even in small-sized or well-differentiated HCC cells, but expression was not detected in AH. It was concluded that PIVKA-II is a useful immunohistochemical marker, even in small-sized or well-differentiated HCC.     

Cytodiagnosis of hepatocellular carcinoma in fine-needle aspirates of the liver: its differentiation from reactive hepatocytes and metastatic adenocarcinoma

Fine-needle aspiration (FNA) cytology is a useful tool for diagnosis of primary malignancies and metastatic lesions of the liver. However, well-differentiated hepatocellular carcinoma (HCC) may resemble benign/reactive hepatocytes, and less differentiated HCC may simulate poorly differentiated adenocarcinoma, leading to difficulties in interpretation of aspirates from liver. To determine the subtle cytomorphological features which can differentiate these lesions, ultrasound-guided FNA smears from 86 cases of liver malignancy were subjected to detailed cytologic assessment. These included 20 cases of HCC, 38 cases of metastatic adenocarcinoma, and 28 cases of benign/reactive hepatocytes. The important features for separating HCC or well-differentiated HCC from benign/reactive hepatocytes were excessive cellularity, trabecular pattern vs. thin cords of hepatocytes, nuclear pleomorphism, atypical stripped nuclei, and macronucleoli (P < 0.001 to < 0.0001). The most significant features for differentiating HCC from metastatic adenocarcinoma were trabecular growth pattern, hepatocytic cells vs. columnar/cuboidal cells, eosinophilic granular cytoplasm, lipid vacuoles, bile pigments, and atypical stripped nuclei (P < 0.001 to < 0.0001). The cytomorphological features which may distinguish poorly differentiated HCC from poorly differentiated adenocarcinoma were polygonal (hepatocytic) cells, eosinophilic granular cytoplasm and lipid vacuoles in HCC, and columnar/cuboidal cells and acinar/glandular formation in adenocarcinoma (P < 0.05 to < 0.001). Diagn. Cytopathol. 1999;21:370-377.     

Survivin在肝细胞癌中的表达特点及其与预后的关系

目的: 研究原发性肝细胞癌组织中Survivin的表达及其与肝癌细胞的生物学行为及预后的关系。 方法: 应用免疫组织化学染色对83例肝癌及相应癌旁组织中Survivin蛋白的表达情况进行检测，应用半定量RT-PCR技术对11例肝癌及相应癌旁组织中Survivin mRNA的表达情况进行检测，结合临床病理资料分析。 结果: 肝癌及癌旁组织中Survivin蛋白的表达率分别为63.9%(53例)和39.6%(21例)。肝癌组织中，41.5%(22例)表达于肝癌细胞核，45.3%(24例)表达于肝癌细胞浆，13.2%(7例)在胞核、胞浆中均有表达。统计学分析表明，Survivin蛋白阳性反应定位于胞核的病例，其肿瘤的包膜侵犯率和转移率更高 (P<0.05)；且术后生存期<2年的肝癌组中的核表达率显著高于术后生存期≥2年者(P<0.01)。RT-PCR结果显示，11例肝癌组织均表达Survivin mRNA，而癌旁组织只有45.5%(5例)表达。 结论: Survivin在肝癌中呈现高表达，其核表达与肝癌的恶性生物学行为及预后密切相关。     

Tumorigenic role of YAP in hepatocellular carcinogenesis is involved in SHP2 whose function is different in vitro and in vivo

Yes-associated protein (YAP) is a nuclear effector of the cell-density sensing Hippo pathway and interacts with Src homology phosphotyrosine phosphatase 2 (SHP2), which controls cell proliferation and survival. The tumor promoting/suppressing activities of YAP and SHP2 during liver tumorigenesis remain controversial. This study aimed to investigate the tumorigenic roles of YAP and SHP2 in hepatocellular carcinogenesis. Cell density associated subcellular distributions of YAP and SHP2 in normal human hepatocytes (THLE-2) and hepatocellular carcinoma (HCC) cells (SK-Hep1, SNU-182) were investigated by Western blotting and cell block immunohistochemistry. The effects of YAP knockdown on proliferation, migration and invasion were studied using YAP-specific siRNAs. The prognostic significance of YAP and SHP2 expressions was investigated immunohistochemically using a tissue microarray (TMA) from 50 HCC cases. High-cell density decreased the nuclear expression of YAP and SHP2 in normal hepatocytes as compared with low-cell density. However, in HCC cells, nuclear YAP and SHP2 were observed regardless of cell density. Nuclear YAP influenced SHP2 expression and cell proliferation. In particular, YAP knockdown impacted nuclear levels of SHP2 protein in SK-Hep1 cells. In HCC tissues, nuclear YAP expression was elevated and cytoplasmic SHP2 expression was diminished as compared with adjacent non-tumor tissues. Notably, these expressions were found to be significantly associated with poor recurrence-free and overall survival rate in patients with HCC. Consequently, the tumor promoting role of YAP is involved in SHP2 which functions as a tumor promoter in vitro but as a tumor suppressor in vivo. YAP and SHP2 can be unfavorable prognostic markers in HCC.     

Utilization of antihepatocyte clone OCH1E5 (Hep Par 1) in histological evaluation of liver tumors

Diagnosis of hepatocellular carcinoma (HCC) is not always easy on simple hematoxylin and eosin (H&E) stain. The diagnostic problems arise when tumor shows pseudoglandular, pleomorphic or clear cell differentiation. Various tumors markers have been described with varying sensitivity and specificity. Monoclonal antibody Hep Par 1 (OCH1E5) which is specific for hepatocytes offers great help in separation of these tumors. The aim of the present study was to determine utility of Hep Par 1 (OCH1E5) in differentiating HCC from metastatic tumors and cholangiocarcinoma. Total of 62 cases of liver tumors obtained from biopsies, resected or autopsy specimens were included in the study. Slides having representative sections were subjected to immunohistochemistry with monoclonal antibody Hep Par 1 (Dako Corp) using avidin biotin technique with primary antibody dilution of 1:40. Adjacent nontumorous hepatocytes were taken as positive control. Slides were examined by experienced pathologist without any information of clinical or H&E diagnosis. Cases were considered positive for Hep Par 1 if tumor cells showed cytoplasmic brown colored granules. The intensity and distribution (diffuse/ focal) of immunoreactivity was noted. Subsequently immunohistochemistry results were correlated with histology and clinical diagnosis. Hep Par 1 antibody was positive in 26 (42 %) and negative in 36 (58 %) liver tumors. On correlating with H&E sections, out of 26 positive cases, 25 (89.2%) were HCC and one was the case of metastasis of mucin secreting adenocarcinoma. From 36 tumors with negative staining 3 were cases of HCC, 27 metastatic adenocarcinomas and 6 cholangiocarcinomas. Only one case of liver metastasis of mucin secreting adenocarcinoma showed positivity. None of the cases of cholangiocarcinoma showed positivity for Hep Par 1. The three HCCs which did not take up staining for Hep Par 1 were 2 cases of moderately differentiated HCC having pseudoglandular pattern and a case of well differentiated HCC with trabecular arrangement. In 11(44%) cases staining was diffuse while in 14 (56%) it was focal but intense. Hep Par 1 is a useful marker in differentiating HCC from metastaic tumors and cholangiocarcinoma with sensitivity and specificity of 89 % and 97 % respectively and positive predictive value of 96 %. However one should be aware of limitations of immunohistochemistry.     

Cytophotometric DNA analysis of hepatocellular carcinoma with Mallory bodies

Summary In order to clarify the pathological significance of Mallory body (MB) formation in human hepatocellular carcinoma (HCC), cell nuclear deoxyribonucleic acid (DNA) content was measured microspectrophotometrically in 20 autopsied cases of HCC associated with cirrhosis and bearing many MBs. According to the degree of dispersion, the DNA histogram was classified into type I (diploid pattern), type II (hyperploid pattern) and type III (aneuploid pattern). Non-neoplastic hepatocytes of normal livers and of cirrhotic areas of the 20 HCC cases showed generally a diploid pattern (type I). In contrast, MB-positive HCC cells showed more hyperploidy or aneuploidy (type I: 0%; type II: 35%; and type III: 65%) compared with MB-negative HCC cells (type I: 25%; type II: 50%; and type III: 25%). These data suggest that MB formation in HCC is accompanied by a constant change of DNA content of HCC cells, though the causal relation between them is only speculative. Two separate HCC nodules in the same liver, both of which contained many MB-positive cells, showed the same type of DNA histogram pattern, suggesting the possibility that they were of a monoclonal origin and had spread discontinuously in the liver.     

Expression of TLR-2 in hepatocellular carcinoma is associated with tumour proliferation,angiogenesis and Caspase-3 expression

AimsUnlike other Toll-like receptors (TLRs), the role of toll like receptor 2 (TLR-2) in the pathogenesis of chronic liver disease and hepatocellular carcinoma (HCC) is not well studied. We, therefore, set out to investigate the expression of TLR-2 in different chronic liver disease states along with other markers of cell death, cellular proliferation and tissue vascularisationMethods and resultsImmunohistochemistry was performed on liver tissue microarrays comprising hepatitis, cirrhosis and HCC patient samples using antibodies against TLR-2, Ki-67, Caspase-3 and VEGF. This was done in order to characterise receptor expression and translocation, apoptosis, cell proliferation and vascularisation. Cytoplasmic TLR-2 expression was found to be weak in 5/8 normal liver cases, 10/19 hepatitis cases and 8/21 cirrhosis patients. Moderate to strong TLR-2 expression was observed in some cases of hepatitis and cirrhosis. Both, nuclear and cytoplasmic TLR-2 expression was present in HCC with weak intensity in 11/41 cases, and moderate to strong staining in 19/41 cases. Eleven HCC cases were TLR-2 negative. Surprisingly, both cytoplasmic and nuclear TLR-2 expression in HCC were found to significantly correlate with proliferative index (r = 0.24 and 0.37), Caspase-3 expression (r = 0.27 and 0.38) and vascularisation (r = 0.56 and 0.23). Further, nuclear TLR-2 localisation was predominant in HCC, whereas cytoplasmic expression was more prevalent in hepatitis and cirrhosis. Functionally, treatment of HUH7 HCC cells with a TLR-2 agonist induced the expression of cellular proliferation and vascularisation markers CD34 and VEGF.ConclusionsOur results demonstrate a positive correlation between the expression of TLR-2 and other markers of proliferation and vascularisation in HCC which suggests a possible role for TLR-2 in HCC pathogenesis     

Morphometric analysis of hepatocellular carcinoma

Summary In order to characterize the cytological features of highly differentiated hepatocellular carcinoma (HCC), a comparative morphometric study was made by observing 30 cases of HCCs and controls (normal, cirrhotic, and atrophic livers). Among trabecular HCCs, normotrabecular subtype (1–2 cell thick cell plate) usually showed minimal cytological atypism and was categorized as well or highly differentiated HCC. Using an image analyzer, the following 4 parameters were applied to quantitate the hepatocyte changes: mean cell size ( ), mean nuclear size ( ), nucleocytoplasmic (N/C) ratio and a coefficient of variance (CV = index of anisokaryosis). In normotrabecular HCCs, was slightly but significantly reduced when compared with normal and cirrhotic livers (t-test:p<0.005). The value was further reduced in mid- and macrotrabecular HCCs. Normotrabecular HCCs showed almost the same value as normal and cirrhotic livers but displayed significantly a higher N/C value than those of controls (t-test:p<0.001). The N/C ratio became even greater in other types of HCCs. While CV was relatively constant in other HCC groups and controls, it was extremely high in the pleomorphic type of HCC and liver cell dysplasia.The results indicated that a reduction in and increase in N/C ratio, which appear as nuclear crowding in histological specimens, actually occurs in well differentiated HCC. For the histologic diagnosis of well differentiated HCC, it would be very important to examine liver specimens with these observations in mind.