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.

Percutaneous injection of a low-concentration alkaline solution targeting hepatocellular carcinoma.

We developed a percutaneous low-concentration alkali injection therapy (PLAIT) targeting hepatocellular carcinoma (HCC), and compared the necrotic areas in the livers of rats that had received PLAIT [an alkaline solution of sodium hydroxide (NaOH)] with those that had received percutaneous ethanol injection therapy (PEIT) and percutaneous acetic acid injection therapy (PAIT). The necrotic area increased with increasing concentrations of NaOH solution. The survival rate of rats was 100% up to a concentration of 4 N; however, the rate dropped below 80% with concentrations over 5 N. Also, at a concentration of 2 N, the necrotic area increased with increasing quantities from 0.01 ml to 0.1 ml. PLAIT using 0.05 ml of 2 N NaOH was 1.56 times more effective than PEIT using 0.05 ml of 99.5% ethanol, and 63.33% less effective than PAIT using 0.05 ml of 50% acetic acid. However, the survival rate after PAIT was 50%, while that after PLAIT was 100%. Histopathological observation of normal rat livers after injection of 2 N NaOH at a volume of 0.05 ml showed that the tissue necrosis spread radially from the site immediately after injection by PLAIT, but necroses were not found in other organs. We conclude that PLAIT has promise as a new form of local therapy for HCC.

Effects of alcohol consumption on histological changes in chronic hepatitis C: a clinicopathological study.

BACKGROUND: To assess the effects of alcohol on the histological changes in chronic hepatitis type C, we performed histopathological examination on liver biopsy specimens by using a semiquantitative method. METHODS: Subjects were 91 patients with chronic hepatitis type C and 32 with alcoholic liver disease. The patients with chronic hepatitis type C were classified into three groups according to the total amount of alcohol intake: nondrinkers, moderate drinkers, and heavy drinkers. For each patient, we evaluated pathological changes of several items and awarded scores from 0 to 2 points, with severe to moderate scoring 2 points, mild 1, and negative 0; the total score was then compared between groups. The evaluated histological changes included virus-related histological changes (V1, inflammatory cell infiltration; V2, lymphoid follicle formation; and V3, bile duct damage) and alcohol-related changes (A1, perivenular fibrosis; A2, stellate/pericellular fibrosis; and A3, fatty change). RESULTS: The total scores of the hepatitis C virus-related histological changes were significantly lower in patients with alcoholic liver disease (ALD group) (p < 0.05). However, we found no significant difference between the different alcohol intake groups. The total score for alcohol-related histological changes significantly increased in line with increases in total alcoholic intake regardless of the presence or absence of hepatitis type C virus infection (p < 0.05). CONCLUSIONS: The results suggest that both alcoholic-related liver damage and virus-related liver damage have specific features; in a addition, alcohol was found to have little effect on the histological liver damage observed in chronic hepatitis type C.

Clinicopathologic significance of protein induced vitamin K absence or antagonist II and alpha-fetoprotein in hepatocellular carcinoma.

Protein induced vitamin K absence or antagonist II (PIVKA-II) and alpha-fetoprotein (AFP) have been considered useful serum markers of hepatocellular carcinoma (HCC). In this study, we examined the clinicopathologic significance of these tumour markers in patients with HCCs by measuring their serum levels and performing immunohistochemistry. We studied 349 Japanese patients with HCCs. Their serum PIVKA-II and AFP levels were determined by enzyme immunoassay before treatment. We examined the correlations between serum PIVKA-II and AFP levels and tumour size, presence of satellite nodules, histologic HCC grade, and concomitant liver diseases and subjected tumour tissues to immunohistochemical staining to detect PIVKA-II and AFP expression. The serum PIVKA-II levels of patients with poorly differentiated HCCs were significantly higher than those of patients with well and moderately differentiated HCCs (p<0.05) and they were higher in HCC patients with than without satellite nodules. The serum AFP levels were influenced significantly by concomitant liver diseases, but not by the other factors. Immunohistochemical staining revealed the PIVKA-II expression levels of poorly differentiated HCCs were higher than those of well and moderately differentiated HCCs (p<0.05). Some HCC cells were PIVKA-II-positive, others were AFP-positive, and some expressed both. The serum PIVKA-II concentration was a better indicator of HCC than AFP, as it was not influenced by concomitant liver diseases. The presence of PIVKA-II in serum correlated with the presence of satellite nodules and the histologic HCC grade, a result concordant with the immunohistochemical findings.

Lewis Y antigen expression in hepatocellular carcinoma. An immunohistochemical study.

BACKGROUND: The altered expression of the Lewis blood group-related antigens during malignant transformation can be used clinically as a tumor marker or as a prognostic indicator. The Lewis Y (LeY) antigen, which is one of the Type 2 human blood group-related antigens, also is thought to behave as an oncodevelopmental cancer-associated antigen. In this study, the authors examined the association between human LeY antigen expression and the clinicopathologic features of HCC, including its proliferative activity. METHODS: Forty-six histologically confirmed cases of HCC were studied retrospectively. Liver biopsy specimens from the main tumor of each case were obtained under ultrasonic guidance before treatment was initiated. The formalin fixed, paraffin embedded serial sections were immunostained using a modification of the avidin-biotin-peroxidase complex method, with a primary monoclonal antibody (MoAb) directed against the LeY antigen (BM-1/JIMRO). The relationship between LeY antigen expression and the HCC's proliferative activity was analyzed similarly by immunohistochemical methods using a primary MoAb directed against the Ki-67 antigen (MIB 1). In addition, to clarify the relationship between LeY antigen expression and the histologic heterogeneity within HCC, seven cases of surgically resected HCC also were immunostained. RESULTS: The LeY antigen was detected on the membrane and in the cytoplasm of the cancer cells. Of the 46 HCC cases, 20 (43.5%) expressed the LeY antigen in the tumor cells. There was no correlation between LeY antigen expression and the maximum tumor dimension or the Stage. However, the incidence of LeY antigen-positive cases in poorly differentiated HCCs was found to be significantly higher than that in well or moderately differentiated HCCs (P < 0.01). In resected HCC cases, LeY antigen expression within HCC nodules was frequently greater in the less differentiated tumor than in adjacent differentiated tumor. Moreover, the incidence of LeY antigen expression in alpha-fetoprotein (AFP)-positive (AFP > or = 200 ng/ml) HCC cases was significantly higher than that in AFP-negative (AFP < 200 ng/ml) HCC cases (P < 0.05). Furthermore, the mean value of the Ki-67 labeling index in LeY antigen-positive HCC cases (25.2 +/- 11.3%) was significantly higher than that in LeY antigen-negative HCC cases (9.4 +/- 4.1%) (P < 0.001). CONCLUSIONS: These results suggest that LeY antigen expression correlated closely to the dedifferentiation and proliferative activity of HCC.

Ultrasonically guided percutaneous microwave coagulation therapy for small hepatocellular carcinoma.

BACKGROUND: The authors have used percutaneous microwave coagulation therapy (PMCT) as a new percutaneous local treatment for single unresectable hepatocellular carcinoma (HCC) measuring 2 cm or less in greatest dimension (small HCC). PMCT was used to attempt a cure of the disease. In this study, the efficacy of this treatment was assessed. METHODS: PMCT was performed on 18 patients with single small HCC. A microwave electrode (custom-made, 30-cm long by 1.6-mm thick) was inserted percutaneously into the tumor area under ultrasonic guidance. Microwaves at 60 W for 120 seconds were used to irradiate the tumor and surrounding area. RESULTS: After PMCT was administered, various image findings were correlated with tissue necrosis. At the tumor and surrounding area, ultrasonography showed echogenic change, contrast enhancement disappeared on contrast enhanced computed tomography, and magnetic resonance imaging (T2-weighted image) showed decreased intensity in all cases after treatment. Complete necrosis of the tumor area in a specimen obtained from one patient who underwent hepatectomy after PMCT also was confirmed. The treatment reduced levels of the tumor marker, alpha-fetoprotein, which had been high in some patients. Although the follow-up period was short (11-33 months), 17 patients remain alive. Local recurrence in the treated area has not been detected, and no serious side effects or complications have been encountered. CONCLUSIONS: PMCT may be an effective and safe treatment for small HCCs.

Laparoscopic observation of retroperitoneal cystic lymphangioma.

A case of retroperitoneal cystic lymphangioma observed laparoscopically is reported. In a 60 year old asymptomatic male patient, an 8 cm multi-loculated cystic lesion was detected incidentally near the splenic hilum with ultrasonography. Endosonography, computed tomography and magnetic resonance imaging revealed the lesion with thin wall and clear fluid. Laparoscopy showed a thin-walled cyst with smooth surface, and straw-coloured clear fluid was observed through the wall. These findings suggested benign aetiology, and seemed to be characteristic of cystic lymphangioma. The tumour was resected, and microscopic examination showed proliferated lymph channels intespersed by lymph follicles. Diagnosis of cystic lymphangioma was established. Laparoscopy seems a useful pre-operative method.

[Evaluation of the biological malignancy in hepatocellular carcinoma by argyrophilic nucleolar organizer region (AgNOR) staining--morphological study of AgNOR using image analyzer].

Using the Colloid silver staining technique to reveal AgNOR and immunostaining for anti-PCNA monoclonal antibody, 23 resected specimens with hepatocellular carcinoma (HCC, < or = 3.5cm in diameter) were examined. These cases were divided into two groups; Group A [9 cases without vascular invasion and a satellite nodule] and Group B [14 cases with satellite nodules]. Comparison of AgNOR score, the morphological features of AgNOR (the area and roundness factor of AgNOR) and PCNA labeling index between Group A and Group B was made by a image analyzer (SP-500). The AgNOR scores and PCNA labeling indices of HCCs in Group B were significantly higher than those of HCCs in Group A. And a close correlation was shown between AgNOR score and PCNA labeling index. Further more, the area, form, and distribution of AgNORs within the nucleus were also different in the two study groups. In Group A, many AgNORs were regular and medium-sized brown dots (AgNOR-roundness factor; > or = 80%, AgNOR-area; 1.5-4.5 microns 2). But in Group B, AgNORs showed marked variation in size and form. These results suggest that HCCs with multiple, smaller, irregular, and widely dispersed AgNOR in combination with high AgNOR scores have a more aggressive potential. The morphological features of AgNOR may be useful indicators for evaluating the proliferative activity of HCC.

[Vanishing tumor thrombus in IVC after oral administration of UFT and TAE in a patient with HCC; a case report].

A 74-year-old man was diagnosed as hepatocellular carcinoma (HCC) in the area of S6-7 with tumor thrombus continuously developed from hepatic vein to inferior vena cava (IVC). AFP value remained high despite two transcatheter arterial embolizations with adriamycin, mitomycin and Lipiodol (Lp-TAE). The AFP value gradually decreased two weeks after oral administration of UFT. Nine months after, the AFP level was 26.4 ng/ml and the tumor thrombus in the IVC disappeared. The size of the main tumor did not change. This case suggests Lp-TAE and UFT are useful for suppressing the growth of the main tumor and to extinguish the tumor thrombus.

Retroperitoneal haematoma due to ruptured microaneurysm of the pancreaticoduodenal artery.

A case of retroperitoneal haematoma due to a ruptured microaneurysm of the posterior superior pancreaticoduodenal artery in a 61 year old man is described. Ultrasonography and computed tomography revealed cystic masses near the gall-bladder. Selective coeliac angiography disclosed a microaneurysm of the posterior superior pancreaticoduodenal artery. Surgical extirpation of the cystic masses was performed, and the histological finding was an encapsulated old haematoma.

Implantation treatment method of slow release anticancer doxorubicin containing hydroxyapatite (DOX-HAP) complex. A basic study of a new treatment for hepatic cancer.

We performed an experimental study on slow releasing anticancer drug implantation treatment as a new therapy for hepatocellular carcinoma. Hydroxyapatite (HAP) was chosen for the carrier material and doxorubicin hydrochloride (DOX) for anticancer agent. DOX-HAP was produced by adsorbing DOX to porous HAP particles of 1375 +/- 125 microns diameter using the freeze drying method. In vitro experiments showed slow release of the drug resulting in the steady release of DOX from HAP for 1 month duration. In healthy white rabbits with DOX-HAP implantation in the liver, serum DOX was not detectable, and DOX release rate was stable at the implanted region after 7, 14, and 21 days. When DOX-HAP (DOX; 100 mg kg-1) was administered to mice with sarcoma 180, an improved survival rate was observed without acute toxicity. We also found that VX2 liver tumour growth on white rabbit was inhibited by implantation of DOX-HAP, without acute toxicity. We hope that DOX-HAP implantation therapy will open up new avenues for the treatment of hepatoma.

A correlation of argyrophilic nucleolar organizer regions with stages of hepatocellular carcinoma.

BACKGROUND: Recently, the number and morphologic features of argyrophilic nucleolar organizer regions (AgNOR), silver-stained nucleolar organizer regions, were thought to reflect the cellular proliferative activity and the grade of malignancy. In liver diseases, it has been reported that AgNOR scores for hepatocellular carcinomas (HCC) were significantly higher than those for benign and borderline lesions; the scores increased with histologic tumor grade. METHODS: Using the colloid silver staining technique to reveal AgNOR, 64 liver biopsy specimens with HCC were examined of which 14 had Stage I disease, 20 had Stage II, 14 had Stage III, 10 had Stage IVA, and 6 had Stage IVB (by International Union Against Cancer criteria). RESULTS: AgNOR in nuclei were divided into two types. Type 1 (T1-NOR) contained large and medium-sized brown dots with well-defined margins. Nucleoli were included in this type. Type 2 (T2-NOR) had fine black single dots and clusters without well-defined margins outside T1-NOR, but within the nucleus. The size and irregularity of T1-NOR changed with the progress of cancerous stages. The increase in the number of T2-NOR contributed preponderantly to the overall increase in Ag-NOR scores. CONCLUSIONS: These results suggest that HCC with smaller and/or irregular T1-NOR in combination with high T2-NOR scores have a more aggressive potential. AgNOR may be useful indicators for evaluating the progress of HCC.