Tatsuo Katagishi

Side effects of high-dose interferon therapy for chronic hepatitis C

BACKGROUND/AIMS Various side effects have been reported in patients treated with alpha interferon, but their incidence and prognosis remain unknown. METHODS Nine hundred and eighty-seven patients with chronic active hepatitis C received 6 to 10 MU of alpha interferon per day for 2 weeks and 3 times per week for 22 weeks. Autoantibodies, thyroid function tests, and fasting plasma glucose concentrations were evaluated prior to alpha interferon therapy. RESULTS Of the 987 patients, 310 were required reduction in the dose of alpha interferon to 3 MU/day or cessation of alpha interferon therapy because of adverse reactions such as flu-like symptoms, leukopenia, and thrombocytopenia. Of the remaining 677, five developed diabetes mellitus, 12 had hyperthyroidism, and six acquired hypothyroidism. Of the 18 with thyroid disorders, five demonstrated antimicrosomal antibodies before therapy. Forty-four patients revealed high or low concentrations of thyroid stimulating hormone at the end of alpha interferon therapy. Three patients developed interstitial pneumonia, one acquired systemic lupus erythematosus-like syndrome, two had autoimmune hepatitis, two developed rheumatoid arthritis, and one developed autoimmune thrombocytopenic purpura. No patients had a history of an autoimmune disorder. One patient experienced sudden hearing impairment and one had retinal detachment. Melena was seen in three patients; two of these cases were compatible with ischemic colitis. Symptoms of depression were seen in 23 patients, and one patient manifested memory loss. CONCLUSION High-dose alpha interferon therapy induces various adverse effects. Most of the side effects cannot be predicted, but are reversible.

Premature telomere shortening and impaired regenerative response in hepatocytes of individuals with NAFLD

Abstract: Aims: The risk factors associated with poor prognosis of nonalcoholic fatty liver disease (NAFLD) are not fully understood. Our aim was to assess the role of progressive hepatocellular telomere shortening in the clinical course of NAFLD.

The copper chelator trientine has an antiangiogenic effect against hepatocellular carcinoma, possibly through inhibition of interleukin‐8 production

Recent studies have revealed that copper is an important cofactor for several angiogenic agents. We examined the antiangiogenic effect against hepatocellular carcinoma (HCC) of the copper chelator trientine, especially focusing on the relationship between copper and interleukin‐8 (IL‐8), a potent angiogenic factor produced by hepatoma cells. HuH‐7 hepatoma cells were transplanted into nude mice and the growth of xenograft tumors was compared to and without administration of trientine. Using the resected tumor, microvessel density, apoptotic potential and proliferative activity were analyzed histologically and IL‐8 mRNA was semiquantified by RT‐PCR. In addition, HuH‐7 cells were cultured in control medium, medium supplemented with copper, medium supplemented with trientine and medium supplemented with both copper and trientine. Human IL‐8 levels were measured in the supernatants by ELISA. Using the extracts from cultured cells, IL‐8 mRNA was semiquantified by RT‐PCR. Trientine suppressed the growth of xenograft tumors significantly. Histologically, apoptotic potential was increased significantly and microvessel density, decreased. The production of IL‐8 from the tumor was suppressed by trientine. In vitro, IL‐8 production by HuH‐7 cells in copper‐containing medium was significantly greater than that in copper‐free medium, and this effect was weakened when trientine was added. However, no significant change was apparent when trientine was added to the medium alone. In conclusion, the chelating effect of trientine prevented copper from functioning as a cofactor in angiogenesis, which resulted in reduced IL‐8 production from HuH‐7 cells.

Role of cell-cycle turnover and oxidative stress in telomere shortening and cellular senescence in patients with chronic hepatitis C.

Background:  In addition to the telomere shortening that occurs with cell division, oxidative stress can damage or shorten telomeres and induce a condition termed premature senescence, possibly before telomeres become critically short. We investigated the effects of cell‐cycle turnover and oxidative stress on cellular senescence in hepatitis C virus (HCV)‐related chronic liver injury.

Centrosome aberration accompanied with p53 mutation can induce genetic instability in hepatocellular carcinoma.

Centrosome duplication is controlled in a cell cycle-specific manner and occurs once every cell cycle, thereby ensuring the balanced segregation of chromosomes during the mitotic phase. Numerical or structural abnormalities can arise in the centrosomes of malignant cells. Under defective cell cycle checkpoint systems, cancer cells with abnormal centrosomes can survive and re-enter the cell cycle, promoting unbalanced chromosome segregation and genetic instability. We investigated the centrosome aberrations in 33 patients diagnosed with hepatocellular carcinoma (HCC), using fluorescent pericentrin immunostaining. We also studied the p53 mutation, proliferative activity, and DNA ploidy in these cases. In normal hepatocytes, one centrosome was identified per cell as a round dot, usually in the vicinity of the nuclear membrane. However, in cancer cells from HCC tissue, several patterns of centrosome abnormalities occurred, including supernumerary centrosomes and centrosomes with an abnormal shape and size. Although the frequency of abnormal centrosomes in each tissue was relatively low compared with previous reports in other cancers, nevertheless, centrosome aberration was found in 30 out of 33 HCC tissues. The percentage of tumor cells with abnormal centrosomes was significantly higher in the nondiploid tumors (15.8±15.9‰) than in the diploid tumors (5.4±5.1‰) (P<0.05), and tended to be higher in the tumors with p53 mutation (11.6±13.1‰) than in those with wild-type p53 (5.6±6.8‰). Furthermore, 82% of nondiploid tumors exhibited p53 mutation, whereas only 41% of diploid tumors showed p53 mutation. The percentage of tumor cells with centrosome abnormalities were not related to tumor stage, size or proliferative activity. Therefore, our results indicate that hepatic cancer cells, under centrosome aberration and a defective checkpoint system possibly caused by p53 mutation, have the potential for genetic instability and aggressive behavior. This potential effect occurs irrespective of the tumor size or stage.

In vivo expression patterns of survivin and its splicing variants in chronic liver disease and hepatocellular carcinoma.

Abstract: Aim: Our aim was to clarify the significance of expression levels and post‐transcriptional splicing patterns of survivin during multistep hepatocarcinogenesis and tumor progression.

Oxidative stress may enhance the malignant potential of human hepatocellular carcinoma by telomerase activation

Background/Aims: Continuous oxidative stress (OS) plays an important role in the progression of chronic liver diseases and hepatocarcinogenesis through telomere shortening in hepatocytes. However, it has not been established how the OS influences the progression of human hepatocellular carcinomas (HCCs). We examined the correlations of OS with telomere length of cancer cells, telomerase activity and other clinicopathological factors in 68 HCCs.

Nuclear size measurement is a simple method for the assessment of hepatocellular aging in non-alcoholic fatty liver disease: Comparison with telomere-specific quantitative FISH and p21 immunohistochemistry

Telomere‐specific quantitative fluorescent in situ hybridization (Q‐FISH) accurately evaluates hepatocellular aging on histological sections, but it requires appropriate tissue processing. To establish a more simple method for the assessment of hepatocellular aging, the usefulness of nuclear size measurement was clarified using biopsy liver samples from 64 patients with non‐alcoholic fatty liver disease (NAFLD), a model for oxidative stress‐associated hepatocellular aging, and 11 control individuals. Relative telomere intensity (RTI) was measured on Q‐FISH, and the relative nuclear size (RNS) was calculated as the average nuclear size of the hepatocytes divided by that of lymphocytes. In normal individuals and NAFLD patients, the RTI and RNS were negatively correlated. The degree of nuclear enlargement in NAFLD patients was larger than that in normal individuals with the same telomere length, possibly reflecting telomere‐independent senescence. In NAFLD patients with RNS >2.0, the regenerative responses, indicated by the ratio of Ki‐67‐positive index to serum alanine aminotransferase level, were significantly reduced. The RNS positively correlated with the p21 expression, another marker of senescence. This all indicates that nuclear enlargement progresses in parallel with reduced regenerative responses, telomere shortening, and p21 upregulation. Nuclear size measurement is an effective method for estimation of hepatocellular aging.

Cytogenetic analyses of hepatocellular carcinoma by in situ hybridization with a chromosome-specific DNA probe

Numerical chromosome analysis has been established in solid tumors by using in situ hybridization (ISH) with a chromosome‐specific probe. We analyzed human hepatocellular carcinoma (HCC) by ISH for chromosome 17 and investigated the correlation of its copy number with histologic malignancy, proliferative activity, p53 mutation, and DNA ploidy.

Identification of apoptotic hepatocytes in situ in rat liver after lead nitrate administration

Apoptosis plays a major role in the regression of mitogen (lead nitrate)-induced hepatic hyperplasia. We compared the in situ end-labeling (ISEL) technique with the conventional detection of apoptotic bodies in this process. In hematoxylin and eosin (H&E) sections, apoptosis is usually recognizable by the presence of apoptotic bodies (apoptosis phase 2). Although the early phase of apoptosis (apoptosis phase 1) can be detected as a prekaryorrhectic appearance in H&E sections, it is difficult to detect and is easily overlooked. On the other hand, ISEL presents intense staining mainly in phase 1 and weak or negative staining in phase 2. Thus, simultaneous investigation by these two methods in two serial sections is the most reliable way to calculate the incidence of apoptosis and gives us precise information on the stages of apoptosis in situ. Since the colorized signals of ISEL are much easier to detect than apoptotic bodies in H&E sections, ISEL is particularly useful for liver tissues, where the incidence of apoptosis is low.