Arterial elements and perisinusoidal cells in borderline hepatocellular nodules and small hepatocellular carcinomas

Borderline hepatocellular nodule in the human cirrhotic liver is considered a preneoplastic lesion of hepatocellular carcinoma (HCC). However, the angiogenetic process and changes in perisinusoidal cells (fat-storing cells or Ito cells) during the borderline nodule-HCC sequence have not been investigated. We have investigated intraparenchymal arterial elements and perisinusoidal cells in normal livers, chronic hepatitis, borderline nodules and small HCC, using an immunohistochemical staining for α-smooth muscle actin. In normal livers, chronic hepatitis, cirrhotic nodules and large regenerative nodules, no or few arterial elements were present in the parenchyma, and α-smooth muscle actin-positive perisinusoidal cells were not increased. In borderline nodules, however, there were many intranodular arterial elements, and perisinusoidal cells were significantly increased. In small HCC, there were much more arterial elements, and perisinusoidal cells were increased further. These data suggest that angiogenesis first occurs in borderline hepatocellular nodules and it gradually proceeds during the nodule to HCC sequence along with an increase in perisinusoidal cells. The demonstration of arterial elements and perisinusoidal cells may be useful for the differential diagnosis of large regenerative nodule, borderline hepatocellular nodule and small HCC.     

Small Liver Masses in Cirrhotic Patients: A Pathological Clue for the Morphogenesis of Human Hepatocellular Carcinoma

Eleven small primary liver lesions detected in four cirrhotic patients are reported in detail. Some lesions showed the histopathological features of macroregenerative nodules (MRN) containing foci of cellular and structural atypia or of hepatocellular carcinoma (HCC). Other lesions showed the histological abnormalities of the early developmental stage of HCC without a background of MRN. Finally one lesion is described with an equivocal background between benign and malignant, containing a focus of carcinoma. All lesions presented suggest the presence of different pathways in the morphogenesis of human HCC.     

‘Atypical adenomatous hyperplasia’ in liver cirrhosis: low-grade hepatocellular carcinoma or borderline lesion?

Adenomatous hyperplasia, defined as a sizable parenchymal nodule in cirrhosis, was examined morphologically. Ninety-seven nodules of adenomatous hyperplasia were obtained from 47 cirrhotic livers and were divided into 'ordinary' (44 nodules) and 'atypical' (53 nodules) types. The former consisted of hepatocytes similar to those of the surrounding liver, and showed regularly distributed portal tracts. The latter type was composed of hepatocytes showing nuclear atypia, relative to the surrounding liver, and showed irregular or sparse portal tracts. Atypical nodules were histologically heterogeneous, possessing areas of normo-trabecular, compact, pseudoglandular and/or scirrhous patterns. Several cytological changes, such as clear cell change, small or large cell change and fatty change, were intermingled variably within a given nodule. Atypical nodules showed expansive and/or replacing growth into the surrounding liver. Atypical hepatocytes also infiltrated into the fibrous septa and portal tracts. Foci of overt hepatocellular carcinoma were found in 11 of the 53 atypical nodules. These findings suggest that ordinary adenomatous hyperplasia may be a large-sized regenerative nodule, while atypical adenomatous hyperplasia may be a hepatocellular neoplasm, a peculiar form of low-grade hepatocellular carcinoma or borderline lesion, in which overt hepatocellular carcinoma is likely to evolve through multiple steps.     

Characterization of hyperplastic foci observed in surgical specimens of hepatocellular carcinoma

By reviewing previous surgical specimens of hepatocellular carcinoma, 17 cases with hyperplastic foci (HPF) characterized by discernible increase in nuclear densities, could be histologically selected. Nuclear densities of HPF and control hepatic parenchyma were assessed quantitatively by counting the nuclear number of hepatic cells, and proliferating cell nuclear antigen labeling index was measured. HPF occurred multifocally, confined within a lobular unit, smoothly merging into surrounding hepatic parenchyma. Nuclear densities of HPF were 1.71 times greater than those of control hepatic parenchyma. The hepatocytes of HPF also showed significantly higher proliferative activities than those of control parenchyma. In addition, noticeable structural distortions, such as focal trabecular thickening or microacinar formation of hepatocytes, were sometimes observed in HPF. However, these HPF seemed to be distinguished from minute de novo hepatocellular carcinoma (HCC) or intrahepatic HCC metastasis, because of paucity of distinctive atypical changes, and intimate correlation with neighboring hepatocytes. Several adjacent HPF were aggregated to form a much larger unit of a hyperplastic area with loss of fibrous septa of liver cirrhosis. It was suggested that grossly detectable large regenerative nodules are produced via fusion of several adjacent HPF.     

Pathologic features of small hepatocellular carcinoma

Twenty-one nodules of small hepatocellular carcinoma (HCC) were examined. Histologically, the nodules often presented formation of plump trabeculae, marked nuclear atypism, or aggressive growth comprising capsular invasion, vascular invasion, and replacement of adjacent pseudolobules. Aside from these characteristic findings of HCC, it was important to reveal the following features for the diagnosis of well differentiated type of small HCC: variable thickening or distortion of trabecular structure in association with nuclear crowding, acinar formation, selective cytoplasmic accumulation of Mallory bodies, nuclear abnormalities consisting of thickening of nucleolus, hepatic cords in close contact with bile ducts or blood vessels, and hepatocytes growing in a fibrous environment. During the invasive growth, the tumor cells may well be subtly blended with benign hepatocytes, giving rise to a pattern of "mixed cellularity". It is also emphasized that connective tissue septa of pseudolobules could be a route of rapid tumor spreading.     

Histopathological and morphometric analysis of atypical adenomatous hyperplasia of human cirrhotic livers

Atypical adenomatous hyperplasia (AAH) is a hyperplastic parenchymal nodular change in the cirrhotic liver, in which overt hepatocellular carcinoma (HCC) occasionally arises. AAH is defined as a sizable hepatocellular nodule with a variable degree of hepatocellular atypia not regarded as HCC, and is different from ordinary adenomatous hyperplasia in which hepatocellular atypia is absent. In the present study, we attempted to evaluate carcinogenetic processes and to find histological variables which indicate malignant transformation in AAH, using 49 surgically resected or autopsied nodules. AAH frequently showed morphological heterogeneity. Atypical lesions within AAHs were divisible into the following three categories from overall histopathological appearances: malignant (A), equivocal (B), or non-malignant (C) lesions. Analysis of combination of these three lesions, which were frequently intermixed in a given AAH, suggested that B lesions appear subsequent to C lesions, and A lesions finally appear in AAH nodules. Among the 14 histological variables, enlargement, hyperchromasia and irregular contour of nuclei were found to correlate well with A lesions. Increased nuclear density, iron resistance, reduction of reticulin fibres, clear cell change, sinusoidal dilatation and presence of abnormal arteries were suggestive of A or B lesions. Nuclear deviation toward the sinusoids, acinar and compact arrangements, fatty change and Mallory's hyaline alone were not useful indicators of A or B lesions. These results indicate that AAH is a preneoplastic or borderline lesion in which overt HCC is likely to evolve through several steps. Although a needle liver biopsy is a useful tool for diagnosis of benign, equivocal and malignant hepatocellular nodular lesions, the needle biopsy specimen should be carefully evaluated by considering the morphological heterogeneity of the AAH and a variable combination of 14 histological variables.     

Diagnostic role of glypican 3 and CD34 for differentiating hepatocellular carcinoma from nonmalignant hepatocellular lesions

Well-differentiated hepatocellular carcinoma (HCC) may be difficult to distinguish from a benign lesion. Glypican 3 (GPC-3) is an oncofetal protein, which has been demonstrated to be up-regulated in HCC. The aim of this study is to evaluate the diagnostic role of combined GPC-3 and CD34 immunoassaying in the distinction between HCC and benign hepatic mimickers. This study was performed on 100 cases of formalin-fixed, paraffin-embedded cases of hepatic focal lesions obtained from the files of pathology laboratory of our university from 2009 to 2012. The following groups were studied: group A (n = 60) (hepatocellular malignant lesions) and group B (n = 40) (Hepatocellular nonmalignant lesions). All cases were stained with GPC-3 and CD34. Sensitivity, specificity, and positive and negative predictive values were calculated for both antibodies. Glypican 3 and complete CD34 staining pattern expression in group A was significantly higher than in group B. The results of costaining showed that, in HCCs, almost all the GPC-3–positive cases had a complete CD34 staining pattern, whereas in the 40 hepatocellular nonmalignant lesions, none stained up with the 2 markers. Therefore, although the sensitivity declined (82%), the specificity and positive predictive value (PPV) of costaining reached 100% and were greater than that observed for single staining with GPC-3 (specificity, 92.5%; PPV, 94.3%) or CD34 (specificity, 97.5%; PPV, 98.3%). Our data demonstrate that GPC-3 and CD34 costaining has better diagnostic value for differentiating HCC from nonmalignant hepatocellular lesions than does single staining.     

Comparative immunohistochemical investigation of rat and human hepatocellular carcinomas

Abstract

Previously identified comparable morphological features of human and rat hepatoproliferative lesions were identified, including hepatocellular carcinomas (HCCs). In this study we identified similarities and differences in immunohistochemical (IHC) detection of some key proteins important in carcinogenesis that may link pathogenesis pathways of human and rat HCC. The comparative features of six IHC markers (Ki-67, beta-catenin, CD34, glutamine synthetase (GS), c-myc, and transforming growth factor alpha (TGF-alpha)), previously shown to be positive or altered in rodent and human HCC when compared to normal hepatocytes, were investigated. Glutamine synthetase (a hepatocellular enzyme) was strongly positive in 5/5 (100%) human and 4/5 (80%) rat HCCs examined. CD34 (an endothelial marker) and Ki-67 (a cell proliferation marker) were consistently positive in five human HCCs (5/5 for both markers) but weakly positive in only 2/5 and 3/5 rat HCCs, respectively. Beta-catenin, c-myc, and TGF-alpha were infrequently altered, with immunopositivity found in only one or two of either rat or human HCCs (a total of five each examined; beta-catenin: 1/5 rat samples, c-myc: 2/5 human samples, TGF-alpha: 1/5 rat samples positive). It was concluded that Ki-67, CD34, and GS, are most likely to be useful in studying the pathogenesis of HCC in rats and humans.     

Diagnostic histopathology of hepatocellular carcinoma: A case-based review

Hepatocellular carcinoma (HCC) is the most common primary hepatic malignant neoplasm in adults. Clinically, it tends to present late, and its prognosis historically has been poor, due to limited treatment options at the more advanced stage. A multidisciplinary approach for surveillance and early diagnosis of hepatocellular carcinoma in high-risk populations, combined with advances in surgical and other forms of ablative or chemotherapy, has greatly improved outcomes for patients with this tumor. Liver biopsies for HCC are becoming rare, being performed only for lesions that are atypical on imaging, to avoid the risk of bleeding or of inadvertently seeding the needle tract with HCC. HCC, a tumor of varied appearance, can be diagnostically challenging, particularly on a liver biopsy, due to sampling errors. Although HCC typically occurs on a background of cirrhosis, or advanced fibrosis, it may also arise in the noncirrhotic liver. Well-differentiated HCC may be hard to distinguish from benign lesions in the noncirrhotic liver and premalignant lesions in the cirrhotic liver, whereas a poorly-differentiated neoplasm may be difficult to tell apart from metastases or primary intrahepatic cholangiocarcinoma. Additionally, variant forms of HCC may mimic other neoplasms. This case-based review discusses typical HCC, the increasingly recognized steatohepatitic variant of HCC, the rare scirrhous variant of HCC and the extremely rare and clinically challenging cirrhotomimetic HCC.     

Expression of both hepatocellular carcinoma and cholangiocarcinoma phenotypes in hepatocellular carcinoma and cholangiocarcinoma components in combined hepatocellular and cholangiocarcinoma

Expression of phenotype markers of hepatocellular carcinoma (HCC) and cholangiocarcinoma (CC) in HCC and CC components of 20 combined hepatocellular and cholangiocarcinomas (CHCs) of the liver was investigated immunohistochemically. Both HCC and CC components of all CHCs expressed at least one of the CC phenotype markers [cytokeratin (CK)-7, CK-19, and carbohydrate (CA) 19-9]. HCC components in 90% of CHCs and CC components in 95% of CHCs expressed at least one of these CC phenotype markers in more than 40% of cancer cells. HCC components in all CHCs expressed at least one of the HCC phenotype markers [hepatocyte antigen (HA), α-fetoprotein (AFP), and canalicular carcinoembryonic antigen]. HCC components in 90% of CHCs and CC components in 75% of CHCs expressed HA, AFP, or both. HCC components in 75% of CHCs and CC components in 60% of CHCs expressed HA, AFP, or both in more than 10% of cancer cells. The present results show that both HCC and CC components of most of the CHCs expressed both HCC and CC phenotypes, supporting the hypothesis that CHC originates from a hepatic progenitor cell capable of differentiating into hepatocytes and cholangiocytes.     

Vascular endothelial growth factor, its receptor Flk-1, and hypoxia inducible factor-1alpha are involved in malignant transformation in dysplastic nodules of the liver

Dysplastic nodules (DNs) are regarded as a premalignant lesion of hepatocellular carcinoma (HCC). Sinusoidal capillarization and unpaired arteries are reported in HCC and also to a lesser degree in DN. However, the mechanism and significance of these vascular alterations remain unclear. In this study, these vascular changes were examined with respect to vascular endothelial growth factor (VEGF) and its receptors (Flt-1 and Flk-1), hypoxia inducible factor-1alpha (HIF-1alpha), and CD34, by using 20 surgically resected cases of DNs and 36 cases of conventional HCC. The expression of these molecules was examined immunohistochemically. Although sinusoidal capillarization characterized by CD34 expression was found diffusely in HCC, such changes were found mainly in the areas around the portal tracts and also in other areas in DNs (focal in 6 cases, zonal in 7 cases, and extensive distribution in 7 cases). These capillarized areas were frequently associated with unpaired arteries, and the infiltration of neoplastic hepatocytes into the portal tracts and loss of reticulin fibers in DNs, particularly those with a zonal and extensive distribution. VEGF was diffusely expressed in neoplastic hepatocytes of DNs and HCC. Interestingly, Flk-1 and HIF-1alpha were mostly expressed in endothelial cells and neoplastic hepatocytes in the capillarized areas around portal tracts in DNs, respectively. In conclusion, the capillarized areas with increased numbers of unpaired arteries in DNs may represent an early malignant transformation. Increased expression of Flk-1 and HIF-1alpha associated with VEGF may be involved in sinusoidal capillarization and the increased numbers of unpaired arteries in these areas.     

Differential expression of beta-galactoside alpha2,6 sialyltransferase and sialoglycans in normal and cirrhotic liver and hepatocellular carcinoma

SUMMARY: Sialyltransferases sialylate plasma glycoproteins in hepatocytes and may (as hepatic key enzymes) constitute markers for liver diseases. We examined expression of the prevalent alpha2,6 sialyltransferase (ST6Gal I) and sialoglycans in normal liver, cirrhotic liver, and hepatocellular carcinoma (HCC) using a new ST6Gal I-specific mAb and recombinant fusion proteins of CD22 and sialoadhesin recognizing alpha2,6- or alpha2,3-sialylated glycans in immunohistology and flow cytometry. In normal and cirrhotic liver, ST6Gal I and sialoglycans were localized in the Golgi region of hepatocytes surrounding the bile canaliculi and along the bile canaliculi, respectively. Sialoglycans were additionally recognized in Kupffer cells, bile ducts, endothelial cells, and oval cells. Well-differentiated and moderately differentiated HCC showed Golgi and diffuse cytoplasmic staining of ST6Gal I and sialoglycans, whereas the cytoplasmic staining for ST6Gal I and sialoglycans was decreased or even absent in poorly differentiated HCC. Detection of sialoglycans by the recombinant fusion proteins in Western blots of cell lysates derived from cell lines revealed two major double bands of sialoglycoproteins at 65 and 120 kDa for hepatocytes, three major bands at 54, 49, and 44 kDa for colonic epithelial cells, and one band at 60 kDa for endothelial cells. Our results describe the expression patterns of ST6Gal I and sialoglycans in various liver tissues and demonstrate an altered expression of these structures between benign and malignant hepatocellular lesions.     

Glypican-3 as a potential differential diagnosis marker for hepatocellular carcinoma: a tissue microarray-based study

The differential diagnosis between hepatocellular carcinoma (HCC) and benign hepatic lesions is still difficult and new biochemical markers for HCC are required. The aim of this study was to assess the differential diagnostic value of glypican-3 (GPC3) immunostaining in HCC patients. 147 cases of surgically excised HCC tissues, 94 cases from needle biopsies, and tissue microarrays were used for this study. The tissue microarrays contained 449 specimens including: 115 HCC, 25 intrahepatic cholangiocellular carcinoma, 29 lung adenocarcinoma, 23 squamous cell lung carcinoma, 53 ovary adenocarcinoma, 44 renal cell carcinoma, 30 prostate acinar adenocarcinoma, 42 breast carcinoma, 41 gastric carcinoma and 47 colorectal carcinoma. The immunolocalization of GPC3 was measured using immunohistochemical staining. Among 147 surgically excised HCC samples, 87.1% (128/147) were GPC3 positive. No GPC3 expression, however, was observed in paracarcinomatous and cirrhotic tissues. In needle biopsy tissues, GPC3 was positively expressed in 81.9% (77/94). Among tissue microassays, HCCs showed positive GPC3 expression in 55.7% (64/115), while 9.6% (5/52) of lung carcinoma and 5.7% (3/53) of ovary adenocarcinoma also were positively stained. The other tumor types showed negative GPC3 expression. In conclusion, our results show that GPC3 is specifically overexpressed in HCC tissue and may be regarded as a potential marker for differential diagnostic hepatocellular carcinoma.     

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.     

Morphology of foci of altered hepatocytes and naturally-occurring hepatocellular tumors in F344 rats

Summary The morphology of liver tumors of F344 rats used as controls in carcinogenesis bioassays were studied. Foci of cellular alteration composed of hepatocytes with basophilic cytoplasm were found commonly in F344 rats, 2 years of age. Eosinophilic and vacuolated foci were considerably less common. The morphology of 67 nodular hepatic lesions indicated that 54 were neoplastic nodules and 13 hepatocellular carcinomas. The majority of these tumors were composed of basophilic hepatocytes. Some of the carcinomas appeared to arise within neoplastic nodules. No tumors metastasized.     

Diagnostic utility of immunohistochemistry in hepatocellular carcinoma, its variants and their mimics.

Hepatocellular carcinoma (HCC) is known for its histomorphologic heterogeneity. Immunohistochemistry (IHC) can help in the comparative morphologic evaluation of HCC, its variants and their mimics. Some of these diagnostic challenges can be attributed to (i) the variety of neoplasms that can arise from the hepatic stem cell lineage; (ii) the spectrum of well-differentiated hepatocellular nodular lesions; (iii) the liver being a target for metastases with some of these histologic entities mimicking variants of HCC or actually arising in the liver; and (iv) the limitations of serum alpha-fetoprotein (AFP). The role of IHC is in the distinction of benign hepatocellular nodules from reactive hepatocytes; WD-HCC from benign hepatocellular nodules; poorly differentiated HCC from cholangiocarcinoma and metastases; and determination of histogenesis of malignant tumor; and of primary site of origin of malignant tumor. A panel of antibodies has more discriminant value. AFP expression usually indicates malignancy in a hepatocellular nodule and hepatocytic histogenesis of a malignancy. Polyclonal carcinoembryonic antigen (pCEA) and CD10 stain bile canaliculi in better-differentiated HCC. HepPar1 is generally accepted as a hepatocytic marker. However, not all HCC stain uniformly and not all HepPar1-positive tumors are of hepatocytic origin or arise in the liver. Mature hepatocytes and hepatocellular nodules stain with CAM 5.2, CK 8, and 18 but not with CK 7, 19, 20, or AE1/AE3. Biliary epithelium expresses CK 7 and 19. CD 34 highlights sinusoidal capillarization. AFP, pCEA/CD10, and CD34 are useful for ascertainment of malignancy in hepatocellular nodules; HepPar1 and cytokeratins to be included if histogenesis is the issue. IHC results should be interpreted in the larger context of the case.     

Glypican-3 expression in hepatocellular tumors: diagnostic value for preneoplastic lesions and hepatocellular carcinomas

Glypican-3 (GPC3), a member of heparan sulfate proteoglycans, plays a role in cell growth, differentiation, and migration. The objectives of this study were to assess the diagnostic value of GPC3 immunostaining in hepatocellular carcinomas (HCCs) and to analyze its expression profile in preneoplastic lesions. Tissue microarrays were built by sampling 54 HCCs and adjacent liver tissues (21 developing from cirrhosis and 33 from normal liver) and 94 cirrhotic macronodules. Fourteen typical liver cell adenomas and 5 with malignant foci were also included. Sections were assessed for GPC3 expression by immunohistochemistry. GPC3 staining was observed in 19 (90%) of 21 HCC cases with cirrhosis and in 18 (64%) of 28 HCC cases with normal liver (P < .01). When staining was positive, it was both membranous and cytoplasmic. Positive staining was observed in 1 case of nonneoplastic adjacent liver. In cases of adenomas, only malignant foci were positive. Among the 94 macronodules, GPC3 immunostaining was noted in 48% (14/29) of high-grade dysplastic or early HCC and in 3% (2/65, P < .001) of benign or low-grade dysplastic macronodules. This study shows that GPC3 is an efficient diagnostic marker of HCC, potentially useful in the differential diagnosis of liver cell adenomas and well-differentiated HCC. Our results also suggest that GPC3 may be considered as an early marker of liver carcinogenesis because it is able to identify some cirrhotic macronodules with malignant potential.     

锌指蛋白281对肝癌细胞增殖的影响

目的:研究锌指蛋白281(zinc finger protein 281,ZNF281)对肝癌细胞增殖的作用。方法:用realtime PCR检测80例健康人群及206例肝癌患者外周血单个核细胞中ZNF281的mRNA表达水平,并分析肝癌患者单个核细胞中ZNF281的表达水平与多个临床病理参数的相关性。采用real-time PCR及Western blot实验分别检测ZNF281在肝癌细胞和永生化肝细胞中的mRNA及蛋白表达水平。利用小干扰RNA靶向沉默Huh-7细胞和SKHep-1细胞中ZNF281的表达,然后用MTS法检测肝癌细胞活力的变化;EdU标记实验检测肝癌细胞增殖能力的变化;平板集落形成实验检测肝癌细胞集落形成能力的变化;软琼脂集落形成实验检测肝癌细胞锚定非依赖生长能力的变化。结果:与正常健康人群相比,肝癌患者的外周血单个核细胞中ZNF281的mRNA表达水平明显增高,且其增高水平与肿瘤的大小、分期以及血管侵袭性存在正相关。肝癌细胞中ZNF281表达水平高于永生化肝细胞(P0.05)。沉默ZNF281可以抑制肝癌细胞的活力,抑制肝癌细胞增殖,DNA合成降低,集落数量减少,锚定非依赖性生长能力减弱(P0.05)。结论:ZNF281可促进肝癌细胞的增殖。     

Impaired autophagy response in human hepatocellular carcinoma

Background

Autophagy is a cellular lysosomal degradation mechanism that has been implicated in chronic liver diseases and hepatocellular carcinoma (HCC). Association of autophagy defect with the development of human HCC has been shown in transgenic mouse model.

Aim

We performed this study to verify whether a defect in autophagy would play a role in human hepatocellular carcinoma (HCC).

Methods

Archival tissue sections of 20 patients with HCC with or without hepatitis C virus (HCV) infection were studied. All slides were immunostained using monoclonal antibodies to p62 and glypican-3 with appropriate positive and negative controls. The expression of p62 and glycican-3 in the HCC and the surrounding non-tumor was semiquantitated. The cytoplasmic staining was graded as negative, weak or strong.

Results

Positive p62 staining was found in 20 out of 20 (100%) HCCs and negative staining was observed in 20 out of 20 non-tumor areas and cirrhotic nodules. Positive glypican-3 staining was found in 70% of HCCs and negative staining was seen in all non-tumor areas. An autophagy defect leading to increased expression of p62 and glypican-3 was also seen in the HCC cell line (Huh-7.5), but not in the primary human hepatocytes. Activation of cellular autophagy in Huh-7.5 cells efficiently cleared p62 and glypican-3 expression and inhibition of autophagy induced the expression of p62 and glypican-3.

Conclusions

This study shows that p62 is increased in HCC compared to the surrounding non-tumorous liver tissue suggesting that human HCCs are autophagy defective. We provide further evidence that glypican-3 expression in HCC may also be related to defective autophagy. Our study indicates that p62 immunostain may represent a novel marker for HCC.     

肝癌组织中ras,c—myc,c—erbB—2和p53的蛋白表达

目的：探讨肝癌的发病机理，方法：应用免疫组化ＳＰ法对４６例肝癌细胞组织及其３８列癌旁中癌基因ｒａｓ，ｃ－ｍｙｃ，ｃ－ｅｒｂＢ－２和抑癌基因ｐ５３基蛋白达作定位研究，结果：在癌组织中的检出率分别为５８．７％，６７．４％，４７．８％和２０．４％，癌旁组织中的检出率分别为３４．２％，４７．４％，４２．１％和７．９％，ｒａｓｐ２１蛋白位于肝细胞和肝细胞的胞浆内ｃ－ｅｒｂＢ－２蛋白位于肝细胞和肝癌细胞的胞浆