Yukifusa Kuni

Development of hepatocellular carcinoma associated with increases in DNA synthesis in the surrounding cirrhosis

The relationship between DNA synthesis activity in hepatocytes from cirrhotic tissue and development of hepatocellular carcinoma was studied in 33 posthepatitic patients with Childs grade A cirrhosis. DNA synthesis activity was measured by a bromodeoxyuridine (a thymidine analogue) labeling index, using the bromodeoxyuridine-antibromodeoxyuridine in vitro method, and the patients were followed up prospectively with frequent liver ultrasonography for 2 years. During the 2-year follow-up, 11 of the 33 cirrhotic patients developed hepatocellular carcinoma; they included 8 of the 15 patients (53%) in the high labeling index (greater than 1.5%) group compared with only 3 of the 18 patients (17%) in the low labeling index (less than 1.5) group (P less than 0.05). Five of the latter 18 subsequently had increased synthesis activity. Of these 20 patients who showed high synthesis activities either initially or subsequently, 10 (50%) developed hepatocellular carcinoma, in contrast to 1 of 13 (8%) with persistently low activities (P less than 0.05). Thus, hepatocellular carcinoma seems to develop or may become detectable when DNA synthesis in the background cirrhosis is increasing or remains high.

Difference in the in vitro uptake of bromodeoxyuridine between liver cirrhosis with and without hepatocellular carcinoma

With the aim of examining increasing deoxyribonucleic acid (DNA) synthesis of liver cirrhotic tissue when hepatocellular carcinoma (HCC) is developing, bromodeoxyuridine labeling indices (BrdU LI) of liver biopsy specimens from 19 control cirrhotic (control LC) patients without HCC, 19 cirrhotic patients with HCC, and from six control subjects were examined using an in vitro labeling technique. The mean BrdU LI ± SD of HCC cancerous portion, HCC non‐cancerous cirrhotic portion, control LC, and of control subjects were 7.2 ± 2.9%, 3.3 ± 1.5%, 2.1 ± 1.7%, and 0.25 ± 0.09%, respectively. Interesting enough, there was a significant difference (P < 0.05) between the non‐cancerous cirrhotic portion and control LC. Although all 17 non‐cancerous cirrhotic portion belonged to the high LI group (> 1.5%), 10 of 19 in the control LC belonged to the low LI group (>1.5%) (P > 0.001). The authors conclude that HCC would develop in the cirrhotics with high DNA synthetic potency.

In vitro uptake of bromodeoxyuridine by human hepatocellular carcinoma and its relation to histopathologic findings and biologic behavior

The in vitro uptake of bromodeoxyuridine (BrdU) by hepatocellular carcinoma (HCC) cells was studied in 30 hepatectomized patients. Labeling of the nuclei by BrdU expressed as labeling index (LI) was 5.6 ± 3.2% (mean ± standard deviation), with a considerable variation from case to case. The mean LI in Grade III to IV cancers (less differentiated, by Edmondson and Steiners classification, 11.1 ± 2.1%) was significantly larger (P < 0.001) than that in Grade I to II cancers (more differentiated, 4.5 ± 2.0%). Capsule formation was found in all 17 patients except one (94%) with a low DNA synthetic HCC (LI < 6.0%) compared with seven of 13 (54%) with a high DNA synthetic HCC (LI ± 6.0%, P < 0.02). The 2‐year survival rate after surgery was significantly higher (P < 0.02), and intrahepatic metastasis was significantly less (P < 0.05) in the former group than in the latter. The BrdU LI of HCC tumors showed a strong correlation with histopathologic findings and the biologic behavior of HCC.

Increased uptake of bromodeoxyuridine by hepatocytes from early stage of primary biliary cirrhosis.

The relationship between DNA synthesis activities of hepatocytes in biopsied specimens and liver volume was studied in various stages of primary biliary cirrhosis using an in vitro bromodeoxyuridine (a thymidine analogue)-anti-bromodeoxyuridine reaction and computed tomography. The mean bromodeoxyuridine (+/- SE) labeling index for 10 patients in an early histological stage (stage I, 4, and stage II, 6, 3.4% +/- 0.4%) of primary biliary cirrhosis was 17 times that for 6 control subjects (0.2% +/- 0.1%, P less than 0.001), and was significantly higher than that for 19 female patients with chronic aggressive hepatitis (0.9% +/- 0.2%, P less than 0.001), 14 compensated cirrhotic patients of viral origin (all female, 1.1% +/- 0.3%, P less than 0.01), and 5 patients with stage III primary biliary cirrhosis (0.5% +/- 0.1%, P less than 0.001). The mean (+/- SE) liver volume in the early stage of primary biliary cirrhosis (1225 +/- 40 cm3) was about 1.5 times that in control subjects (835 +/- 42 cm3, P less than 0.001). These results suggest that liver volume has already become large in the early stage of primary biliary cirrhosis perhaps because of markedly increased DNA synthesis in hepatocytes.

A Survey of Regenerating Capacity in Patchy Liver and in Nodular Liver—with Special Reference to DNA Synthesis Estimated by BrdU Labeling Index—

Abstract: Laparoscopically, patchy liver and nodular liver were considered as features of regeneration of liver cells. In this study, the regenerating capacity of liver cells obtained by a liver biopsy from laparoscopically identified patchy liver and nodular liver was estimated by Bromodeoxvuridine (BrdU)‐anti‐BrdU method. BrdU labeling indices (L. I.), of liver cells in biopsy specimens from 6 normal livers, 12 patchy livers, and from 15 nodular livers were examined using an in‐vitro labeling technique. Liver biopsy specimens obtained by a Tru‐cut needle were immediately incubated for 45 min. in 0.1% BrdU solution in RPMI 1640 at 37°C under a pressure of 3 atmospheres in a mixture of 95% O2 and 5% CO2. Immunohistochemical detection of BrdU was performed by the Avidin‐Biotin‐Peroxidase Complex (ABC) method. The mean BrdU L. I. (±SE) of normal liver, patchy liver, and of nodular liver was 0.25±0.09%, 1.4±0.2%, and 1.7±0.4% respectively. Among the nodular liver, flat shaped ones showed a low level (0.5±0.1%), in contrast to the high level (2.4±0.7%) in the semispherical nodular liver. The BrdU L. I. of both the patchy liver and the nodular liver was significantly higher than that in the normal liver (p<0.001, p<0.01 respectively).

Increased Deoxyribonucleic Acid Synthesis in Primary Biliary Cirrhosis with the Laparoscopical Hallmark of “Reddish patch”

Abstract: The deoxyribonucleic acid (DNA) synthetic potency of hepatocytes in liver biopsy specimens was investgated in: 6 patients with primary biliary cirrhosis (PBC) with a laparoscopical finding of a “reddish patch”, 5 patients with PBC without a “reddish patch”, 10 patients with chronic aggressive hepatitis with a typical “patchy liver” as seen on laparoscopy, 14 patients with cirrhosis of a viral origin with a laparoscopic “nodular liver”, and in 6 patients with gastric cancer without liver involvement (control subjects). The bromodeoxyuridine (BrdU, a thymidine analogue) ‐anti‐BrdU method was used. The mean BrdU L. I. (±SE) of the PBC patients with a “reddish patch”, PBC patients without a “reddish patch”, patients with a “patchy liver”, patients with a “nodular liver” and of the control subjects was 3.5±0.6%, 0.5±0.1%, 1.3±0.3%, 1.1±0.3%, and 0.2±0.1% respectively. The BrdU L.I. of the PBC patients with a “reddish patch” was significantly greater than that of the PBC patients without a “reddish patch” (p>0.001) and greater than that of patients with a “patchy liver” or a “nodular liver” (p>0.001, respectively). It was the greatest in all of the benign liver diseases examined. Thus, the so‐called laparoscopical finding of a “reddish patch” in PBC may represent a marked regeneration of hepatocytes.