Hisashi Matsunaga

Protein-bound polysaccharide PSK inhibits tumor invasiveness by down-regulation of TGF-β1 and MMPs

Transforming growth factor β1 (TGF-β1) and matrix metalloproteinases (MMPs) produced by tumor cells play important roles in tumor invasion. PSK, a protein-bound polysaccharide, is widely used in Japan as an immunopotentiating biological response modifier for cancer patients. In this study, we focused on the effects of PSK on invasiveness, TGF-β1 production, and MMPs expression in two human tumor cell lines, pancreatic cancer cell line (NOR-P1) and gastric cancer cell line (MK-1P3). PSK significantly decreased the invasiveness of both cell lines through Matrigel-coated filters but did not affect cell viability, proliferation, or adhesion. Decreased invasion was associated with the inhibition of TGF-β1, MMP-2, and MMP-9 at both mRNA and protein levels as assessed by reverse transcriptase-polymerase chain reaction, gelatin zymography, and enzyme-linked immunosorbent assay. Antibody against TGF-β1 neutralized the MMP activities of both cell lines. PSK also suppressed the expression of urokinase plasminogen activator (uPA) and uPA receptor but did not change plasminogen activator inhibitor-1 (PAI-1) expression. Western blot analysis showed that PSK reduced uPA protein expression but not PAI-1 expression in the both cell lines. These results indicate that PSK suppresses tumor cell invasiveness through down-regulation of several invasion-related factors including TGF-β1, uPA, MMP-2, and MMP-9.

Potentiation of cytotoxicity of mitomycin C bt a polyacetylenic alcohol, panaxytriol

Polyacetylenic alcohol, panaxytriol, which was isolated fromPanax ginseng C. A. Meyer, has antiproliferative activity against several kinds of tumor cells. In this paper, the effect of panaxytriol on the cytotoxicity of mitomycin C (MMC) against a human gastric carcinoma cell line, MK-1, was investigated. The combination of a subthreshold concentration of MMC and panaxytriol produced a significant cytotoxic effect, which indicates that the effects of panaxytriol and MMC are synergistic. A synergistic effect was observed when MK-1 cells were treated with the mixture of MMC and panaxytriol or treated with MMC followed by panaxytriol. In contrast, when MK-1 cells were exposed to panaxytriol and then to MMC, only an additive effect was induced. With the aim of finding a possible mechanism, the effect of panaxytriol on the accumulation of MMC into the MK-1 cells was examined. Cellular concentration of MMC were measured by highperformance liquid chromatography (HPLC). When MK-1 cells were treated with a mixture of panaxytriol and MMC or first with MMC and then with panaxytriol, the cellular level of MMC was significantly higher than that in MK-1 cells treated with MMC alone, but no significantly increased accumulation was found when MK-1 cells were treated with panaxytriol followed by MMC. These results suggest that synergistic effects of panaxytriol and MMC may be induced by acceleration of the effect of MMC on cellular that the enhanced accumulation of MMC in MK-1 cells treated with panaxytriol can probably be attributed to the decreased fluidity of the cell membrane caused by panaxytriol.

A possible mechanism for the cytotoxicity of a polyacetylenic alcohol, panaxytriol: inhibition of mitochondrial respiration

A polyacetylenic alcohol, panaxytriol, isolated fromPanax ginseng C. A. Meyer inhibits both tumor cell growth into mice. Our preliminary studies indicated that panaxytriol localizes to the mitochondria in human breast carcinoma cells (Breast M25-SF). This study focused on the effects of panaxytriol on mitochondrial structures and function in Breast M25-SF. The results indicate that panaxytriol rapidly inhibits cellular respiration and disrupts cellular energy balance in Breast M25-SF. At concentrations between 11.3 and 180 μM, panaxytriol causes a dose-dependent inhibition of the conversion of the tetrazolium (MTT assay) by mitochondrial dehydrogenase within 2 h. A 1-h treatment with 180 μM panaxytriol causes a significant loss of rhodamine-123 from cells with mitochondria prestained with rhodamine-123 (by flow cytometry). Specific toxic changes were observed by electron microscopy in the mitochondria of Breast M25-SF within 1 h after treatment with more than 180 μM panaxytriol. These data indicate that 180 μM panaxytriol rapidly disrupts cellular energy balance and respiration in Breast M25-SF and suggest that panaxytriol may lower cellular ATP concentrations. After treatment with 180 μM panaxytriol, cellular ATP levels were 40% of those in control cells after 1 h. ATP depletion preceded the loss of cellular viability. Neither ATP depletion nor cytolysis was found in human erythrocytes that have no mitochondria. Thus, ATP depletion resulting from a direct inhibition of mitochondrial respiration is a critical early in the cytotoxicity of panaxytriol.

Novel antiproliferative falcarindiol furanocoumarin ethers from the root of Angelica japonica.

Four novel antiproliferative furanocoumarin ethers of falcarindiol, named japoangelols A (8.5), B (7.2), C (7.4), and D (8.4), were isolated from the root of Angelica japonica together with panaxynol (0.3), falcarindiol (3.2), (9Z)-1,9-heptadecadiene-4,6-diyne-3,8,11-triol (2.2), and 8-acetoxyfalcarinol (3.2). Structures were established from the spectroscopic evidence, and the inhibitory activities (ED50, microgram/ml, shown in the parentheses) were evaluated using the MTT assay.

Oral dimethyl sulfoxide for systemic amyloid A amyloidosis complication in chronic inflammatory disease: a retrospective patient chart review.

BackgroundAmyloid A amyloidosis is an obstinate disease complication in chronic inflammatory disease, and there are few effective therapies. The objective of this study was to investigate the effect of oral dimethyl sulfoxide (DMSO) on amyloid A amyloidosis.MethodsFifteen secondary amyloid A amyloidosis patients (4 men, 11 women; age, 23–70 years) were treated with DMSO between 1995 and 2003. DMSO was administered orally in all patients at a dose of 3–20 g/day. The clinical symptoms together with the renal and gastrointestinal functions were evaluated before and after treatment.ResultsAmong the 15 patients, amyloid A amyloidosis was a complication of rheumatoid arthritis (RA) in 10, of Crohns disease in 4, and of Adult Stills disease in 1. Nine cases mainly involved the kidney, with renal dysfunction and proteinuria, five mainly involved the gastrointestinal tract, with protein-losing gastroenteropathy and intractable diarrhea, and one involved both gastrointestinal and renal amyloidosis. DMSO treatment was successful in 10 (66.7%) of the 15 patients (RA, 6/10; Crohns disease, 4/4; Adult Stills disease, 0/1). Eight weeks of DMSO administration improved the renal function and proteinuria in five out of ten renal amyloidosis patients, but had no effect on those patients with severe and/or advanced renal dysfunction. With regard to gastrointestinal amyloidosis, gastrointestinal symptoms, including diarrhea and protein-losing gastroenteropathy, were improved in six patients. No serious side effects were encountered with the DMSO treatment.ConclusionsOral administration of DMSO is an effective treatment for amyloid A amyloidosis, especially for gastrointestinal involvement and the early stage of renal dysfunction.

Studies on vocal fold injection and changes in pitch associated with alcohol intake

The current study was carried out with particular emphasis on the association between phonetic function tests and alterations in the appearance of the hypopharyngeal and laryngeal mucosa, such as capillary dilatation, edema, and vocal fold injection after alcohol intake. The results demonstrated the occurrence of previously unrecognized pathophysiological changes associated with synchronous phonetic functions in the vocal pathway after alcohol intake. Serum ethanol and aldehyde concentration levels were evaluated hourly for 2.5 h after ingestion of alcohol. When an electronystagmogram showed the typical pattern of alcohol intake, the study was initiated. Occasionally, rhinography was performed on subjects complaining of a stuffy nose after alcohol intake.

Inhibition of interferon-γ-activated nuclear factor-κB by cyclosporin A: a possible mechanism for synergistic induction of apoptosis by interferon-γ and cyclosporin A in gastric carcinoma cells

Abstract We previously reported synergistic induction of apoptosis by IFN-γ plus either cyclosporin A (CsA) or tacrolimus (FK506) in gastric carcinoma cells. In this study, we aimed to elucidate the mechanism for this synergistic induction of apoptosis. IFN-γ plus CsA synergistically induced caspase-3 mediated apoptosis in gastric carcinoma cells. Although IFN-γ induced activation of signal transducer and activator of transcription1 (STAT1) and expression of interferon regulatory factor-1 (IRF-1) mRNA, IFN-γ alone was not able to induce caspase-3 activation and apoptosis. When gastric carcinoma cells were treated with cyclohexamide, a protein synthesis inhibitor, following IFN-γ pretreatment, caspase-3 was activated, and apoptosis was markedly induced. These findings suggest the existence of IFN-γ-induced anti-apoptotic pathway and we evaluated the effect of IFN-γ and CsA on calcium-sensitive nuclear factor-κB (NF-κB) activation. IFN-γ increased intracellular calcium ion concentration ([Ca 2+ ] i ) consisting of a spike and a sustained phase, and the latter was completely abrogated by CsA. Activation of NF-κB occurred in response to IFN-γ, and which was markedly inhibited by either CsA or FK506. NF-κB decoy also enhanced the cytotoxic effect of IFN-γ. These results suggest that IFN-γ may simultaneously induce the STAT1-mediated apoptotic pathway and the anti-apoptotic pathway through calcium-activated NF-κB and that inhibition of the latter by CsA may result in dominance of the apoptosis-inducing pathway.

Expression of interleukin (IL)-12 mRNA in gastric carcinoma specimens: Cellular antitumor immune responses

Several tumor‐related antigen peptides that are recognized by autologous cytolytic T cells (CTL) have been reported. However, most human solid tumors, including gastric carcinoma, are only weakly immunogenic. In this study, we focused on interleukin (IL)‐12 and interferon‐γ (IFN‐γ) as key cytokines for estimating positive cellular immune responses.

Determination of panaxytriol, a new type of tumour growth inhibitor from Panax ginseng, by capillary gas chromatography

Panax ginseng is known for its unique antitumour therapeutical effect’. During a series of studies aimed at isolation of the tumour growth inhibitory substance from Panax ginseng, we found a substance, a polyacetylenic alcohol, which inhibits tumour cell growth in a dose-dependent fashion in vitro ‘y3 Data from infrared (IR), proton and carbon-l 3 nuclear magnetic resonance and high-resolution mass spectra were identical with those of authentic panaxytriol, heptadec-l-ene-4,6-diyne-3,9,10-triol, previously described by several investigators4g5. In order to examine the mechanism of cell growth inhibition caused by panaxytriol, it is important to develop an assay method for panaxytriol. We describe in this paper a gas chromatographic (GC) method for its quantitation by silylation.