Akira Satomi

Significance of increased neutrophils in patients with advanced colorectal cancer

We examined the ratio of neutrophils to lymphocytes (N/L ratio) in the peripheral blood in patients with colorectal cancer. The ability to produce active oxygen and phagocytosis of neutrophils, G-CSF, sIL-2R and IAP (immunosuppressive acidic protein) were also measured. The N/L ratios were significantly higher in the advanced stages of cancer than in normal controls. The ability to produce active oxygen in the terminal stage was 33% lower than in the control group. The G-CSF levels had no relationship with the neutrophil counts. IAP levels increased with cancer stage, and were inversely related to the ability to produce active oxygen. The IAP levels correlated well with the sIL-2R levels and the N/L ratio. These findings suggest that the ability to produce active oxygen, N/L ratio and IAP reflect anticancer mechanisms and that they may be useful when considering treatment or prognosis of patients with advanced stages of cancer.

The proton pump inhibitor inhibits cell growth and induces apoptosis in human hepatoblastoma

PurposeIn normal physiology, a vacuolar-type proton pump (V-ATPase) maintains an intracellular acid microenvironment in lysosome, endosome, and other endomembrane systems. Cancer cells overexpress V-ATPase compared with normal cells, and disturbances of the acid environment are thought to significantly impact the cancer cell infiltration and growth. Bafilomycin A1 (Baf-A1) is a specific inhibitor of the proton-pump inhibitor (PPI) V-ATPase. Neoplastic cells are reportedly more sensitive to Baf-A1 than normal cells, and the difference between the susceptibility to Baf-A1 in normal cells and that in cancer cells may become a target in the cancer therapy. With this in mind, we used cells of hepatoblastoma, the cancer type accounting for 80% of all childhood liver cancers, to investigate the effects of Baf-A1 as an inducer of cancer cell apoptosis and inhibitor of cancer cell reproductionMethods and resultsElectron microscopy showed significant morphological change of the hepatoblastoma cells of the Baf-A1-treated group compared with hepatoblastoma cells of the Baf-A1-free group. The rate of the apoptotic cell increased, and cell reproduction was inhibited. Moreover, the analysis of hepatoblastoma cells using the gene Chip gene expression analysis arrays showed that three of the 27 V-ATPase-related transcripts (ATP6V0D2, ATP6V1B1, and ATP6V0A1) were more weakly expressed in the Baf-A1-treated cells than in the Baf-A1-free cells. In normal human hepatic cells, on the other hand, the inhibition of cell growth of the Baf-A1-treated cells was negligible compared to that of the cells without Baf-A1 treatment. The result of apoptotic cell detection by morphological observations and flow cytometry revealed that Baf-A1 inhibits hepatoblastoma cellular reproduction by inducing apoptosis. On the other hand, the Baf-A1-conferred inhibition of cell growth was negligible in normal human hepatocytesConclusionThe V-ATPase inhibitor Baf-A1 has been proven to selectively inhibit the reproduction and induce the apoptosis of hepatoblastoma cells without adversely influencing normal hepatic cells. With these effects, V-ATPase inhibitors may hold promise as therapeutic agents for hepatoblastoma. Given that three V-ATPase-related genes (ATP6V0D2, ATP6V1B1, and ATP6V0A1) were more weakly expressed in the hepatoblastoma cells of the Baf-A1-treated group than in the Baf-A1-free cells, drug development targeting V-ATPase gene of hepatoblastomas is expected.

Estimation of glycogen levels in human colorectal cancer tissue: relationship with cell cycle and tumor outgrowth

Abstract: In this study, we quantitatively measured glycogen levels in tissue samples obtained from tumors, regions adjacent to tumor, and regions of normal colorectum to determine whether the levels were related to cell cycle and cancer growth. Glycogen levels were analyzed in relation to histopathological factors, (tumor size and stage of disease) and cell cycle progression. The glycogen level was found to be highest in the cancer tissue, lower in normal tissue, and lowest in the adjacent tissue. The difference in glycogen level between the cancer tissue and the other two regions was significant (P < 0.05). There was a negative correlation between glycogen level and tumor size, but it was not significant. The level of glycogen in cancer tissues decreased as the stage of the disease progressed, but a significant difference was not found between stages. There was a negative correlation between the glycogen level and the proliferation index. There was a positive correlation between the glycogen level and the proportion of cancer cells in G1 phase, while there was a negative correlation with S and G2M phases. Glycogen levels were highest in cancers with a high proportion of cells in G1, and decreased with progression to S phase. It may be that glycogen is utilized in the progression to S phase, and the cancer tissues are supplied with glycogen from the tumors themselves as well as their adjacent tissues. Cancer growth may be inhibited by artificial control of the glycogen level in the G1 phase of cancer cells.

Significance of superoxide dismutase (SOD) in human colorectal cancer tissue: Correlation with malignant intensity

The significance of superoxide dismutase (SOD) activity in colorectal cancer tissue was determined from the aspect of the antioxidant defense system. SOD activity and thiobarbituric acid reactive substance were measured in the tumor, in tissues adjacent to the tumor, and in regions that appeared normal, and the results were analyzed in terms of various histopathological factors (stage of disease, depth of invasion, venous invasion, etc.). DNA ploidy pattern and cell proliferation in cancer tissue were also measured, and the results analyzed in relation to SOD activity. SOD activity in cancer tissue was higher than in the other two regions. SOD activity in cancer tissue increased with the progression of stage, and changed with the depth of invasion. There was a significant difference in SOD activity between patients with venous invasion and those in whom this was absent. Stepwise regression analysis suggested that venous invasion was the most significant factor influencing SOD activity. The proliferation index was high in cancer tissue with low SOD activity. The incidence of aneuploidy was high in cancer with high SOD activity, whereas the incidence of diploidy was high in cancer with low SOD activity. These results suggest that elucidation of the antioxidant system in cancer tissue can provide us with a better strategy for cancer treatment.

Serum Soluble Interleukin-2 Receptor in Colorectal Cancer

Preoperative serum soluble interleukin-2 receptor (IL-2R) levels were measured in patients with colorectal cancer, and correlated with various factors as stage, lymph node metastasis, liver metastasis, grade, serum CEA and IAP (immunosuppressive acidic protein). The levels of serum soluble IL-2R in Dukes stage C were significantly higher than in normal control and in Dukes stage A. Serum levels were also significantly higher in patients with lymph node metastasis than in patients without such metastasis, and in IAP positive patients compared to IAP negative patients. Preoperative serum IL-2R levels thus seem to reflect the stage of the disease.

Isolation and identification of cancer stem cells from a side population of a human hepatoblastoma cell line, HuH-6 clone-5

PurposeIt has been thought that the persistence of even a small number of tumor cells in the body may increase each tumor cell in a similar manner and may allow the disease to proceed. However, only a few percent of such tumor cells exist in cancerous tissue. They are called “cancer stem cells (CSCs)”. If an alternative method of annihilating CSCs is found, it will greatly deter relapse and metastasis. We attempted to identify and separate CSCs in hepatoblastoma aiming to develop a new therapy for hepatoblastoma.MethodsThe side population (SP) method was used as an indicator when extracting the CSC candidate group from the hepatoblastoma cells. The SP cells and non-SP cells were studied for tumourigenesis.ResultsAlthough tumors were formed when SP fraction cells were inoculated into mice, tumor formation was not observed in non-SP cells. SP cells had higher tumor formation ability compared to non-SP cells.ConclusionCancer stem-like cells were separated by the SP fraction method from hepatoblastoma cells. The in vivo experiment proved that SP fraction cells inoculated into mice were self-replicated, and the existence of cancer stem-like cells was identified.

Diazotization reaction of nitric oxide trapped by hemoglobin

The present study addresses to determine whether hemoglobin within red blood cells can be utilized as a spin-trap agent for nitric oxide. We demonstrate the diazotization method coupled with a gel filtration chromatography, which is simply due to the separation of nitrosylhemoglobin from nitrite, nitrate or other low molecular nitroso-compounds in biological systems and to the liberation of nitric oxide from nitrosyl heme-complexes in the acidic condition. The amount of nitric oxide can be estimated by the difference of absorbances at 542 nm between diazo-compounds formed by Griess reagent and hemichrome by phosphoric acid. Our results indicate that hemoglobin in red cells as a spin-trap agent would be useful for monitoring nitric oxide in the circulation under the several disease states.

Serum soluble interleukin 2 receptor levels in patients with breast cancer

Preoperative levels of serum soluble interleukin-2 receptor (IL-2R) were examined in 37 patients with breast cancer. We investigated the correlations of serum soluble IL-2R levels with various factors such as stage grouping, lymph node metastasis, distant metastasis, tumor size, histopathological type, estrogen receptor (ER), progesterone receptor (PgR) and CA 153. Serum soluble IL-2R levels were measured with an enzyme-linked immunosorbent assay.Levels of serum soluble IL-2R in the patients with stage III and IV breast cancer were significantly higher than those in the normal controls, and patients with stage I and II breast cancer. Preoperative levels of serum soluble IL-2R in patients with T3 and T4 were also significantly higher than those in patients with T1 and T2. Serum levels of IL-2R in patients with distant metastasis were also significantly higher than those in patients without distant metastasis. Moreover, serum levels of soluble IL-2R in patients with higher CA 15-3 were significantly higher than those in patients with normal CA 15-3 levels.We conclude that preoperative serum soluble IL-2R levels in patients with breast cancer may be a valuable parameter, especially in evaluating whether they have distant metastasis or not.

Applications of Fourier transform infrared spectroscopy, Fourier transform infrared microscopy and near-infrared spectroscopy to cancer research

Glycogen levels in human lung and colorectal cancerous tissues were measured by the Fourier transform (FT-IR) spectroscopic method. Reliability of this method was confirmed by chemical analyses of the same tissues used for the FT-IR spectroscopic measurements, suggesting that this spectroscopic method has a high specificity and sensitivity in discriminating human cancerous tissues from noncancerous tissues. The glycogen levels in the tissues were compared with the clinical, histological and histopathological factors of the cancer, demonstrating that glycogen is a critical factor in understanding the biological nature of neoplastic diseases. Furthermore, direct measurement of a very small amount of tissue by a FT-IR microscope suggested that it could be used as a diagnostic instrument for various tissue samples obtained via a fine needle biopsy procedure. The progressive alterations in rat mammary gland tumors were investigated by a near-infrared (NIR) spectrometer with a fiber optic probe. A lipid band due to the first overtone of n-alkane was used to quantitatively evaluate malignant changes in the tumors. NIR spectroscopy may offer the potential for non‒invasive, in vivo diagnosis of human cancers.

Tissue superoxide dismutase (SOD) activity and immunohistochemical staining in acute appendicitis: Correlation with degree of inflammation

The mechanism of progression of appendicitis has not been clarified. We esamined tissue superoxide dismutase (SOD) activity, thiobarbituric acid reactive substance (TBARS), and the localization of Cu, Zn-SOD in 56 inflamed appendices in relation to histopathological classification. There was a significant difference in SOD activity between catarrhal appendix and phlegmonous and gangrenous appendix (2.3±0.1 vs 5.0±0.2 and 4.6±0.6 units/mg protein, respectivelyP<0.05). TBARS value was highest in gangrenous appendix, being significantly different from the levels in the other two types (0.47±0.40 vs 0.19±0.01 n mol/mg protein, in catarrhal and 0.20±0.02, in phlegmonous appendixP<0.05). Positive staining for Cu, Zn-SOD was demonstrated in 64% of catarrhal appendices, 96% of phlegmonous appendices, and 75% of gangrenous appendices, and intense positive staining was recognized in 9%, 28%, and 40% of these appendices, respectively. These results indicated that active oxygen influences the degree of inflammation in phlegmonous and gangrenous appendicitis. Gangrenous appendicitis and the other two types of appendicits seemed to be different entities.