Shigeo Matsuyama

Expression of Hypoxia-Inducible Factor-1α Protein Predicts Survival in Patients with Transitional Cell Carcinoma of the Upper Urinary Tract

Hypoxia-inducible factor-1 (HIF-1), identified as one of the transcription factors, has been found to play an essential role in oxygen homeostasis. HIF-1 is a heterodimer composed of HIF-1α and HIF-1β. Increased levels of HIF-1α have been reported during the carcinogenesis and progress of several tumors. We investigated the prognostic importance of HIF-1α expression in transitional cell carcinoma of the upper urinary tract. In 127 cases of transitional cell carcinoma of the upper urinary tract, we examined its expression (using immunohistochemistry and in situ hybridization), and also its relation to the expression of p53 oncoprotein, as well as to proliferating cell nuclear antigen (PCNA) immunoreactivity, microvessel density, clinicopathologic parameters, and clinical outcome. A positive expression of HIF-1α protein was recognized in 55.1% of samples, the expression being apparent within the nucleus in tumor cells. HIF-1α protein expression correlated with grade, growth pattern, p53 oncoprotein expression, and PCNA index, but not with stage. Furthermore, a significant correlation was found between HIF-1α protein expression and both overall and disease-free survival rates in the univariate and multivariate analyses (in all tumors and in invasive tumors). A positive expression of HIF-1α mRNA was recognized in 69.6% of 125 samples which were available, the expression being apparent within the cytoplasm in tumor cells. The positive expression of HIF-1α mRNA by in situ hybridization correlated significantly with HIF-1α protein expression by immunohistochemistry. HIF-1α mRNA expression only correlated with pattern of growth (P = 0.0078). In conclusion, the detection of HIF-1α protein would seem to be of value in informing the prognosis of transitional cell carcinoma of the upper urinary tract.

Joint fluid carboxy-terminal type II procollagen peptide as a marker of cartilage collagen biosynthesis

Joint fluid levels of carboxy-terminal type II procollagen peptide (pCOL II-C) were measured in osteoarthritis, rheumatoid arthritis and traumatic arthritis by a newly developed one-step enzyme immunoassay (EIA). The detection limit of the new method was as low as 0.2 ng/ml. The levels of pCOL II-C were significantly (P < 0.001) higher in osteoarthritis and traumatic arthritis than in rheumatoid arthritis. In osteoarthritis, pCOL II-C levels were higher in moderately afflicted patients. Since type II collagen is a unique component of cartilage, pCOL II-C levels in joint fluids could reflect the synthetic activity of type II collagen of chondrocytes in the diseased joint and therefore could be utilized as a simple marker of type II collagen synthesis in articular cartilage in joint diseases.

Gene transfer of human hepatocyte growth factor into rat skin wounds mediated by liposomes coated with the sendai virus (hemagglutinating virus of Japan).

Hepatocyte growth factor (HGF) regulates cell growth, cell motility, and morphogenesis in various types of cells, including epithelial and endothelial cells, indicating that it probably promotes epithelial repair and neovascularization during wound healing. To better understand the effects of HGF on wound healing, we performed human HGF-gene transfer into skin wounds in rats. The rat HGF mRNA levels, and human and rat HGF protein concentrations in the wounds in HGF gene-transfer rats were significantly elevated at 3 days, 3 to 14 days, and 3 and 14 days after gene transfer, respectively. An expression of human HGF mRNA and protein was revealed in squamous cells in the epidermis, in endothelial cells and smooth muscle cells in blood vessels, and in fibroblasts in granulation tissues at 3, 7, and 14 days after gene transfer in HGF gene-transfer rats. The wound lesion area in HGF gene-transfer rats was significantly less than that in control rats from 3 to 7 days after gene transfer. The re-epithelialization rate, microvessel counts in granulation tissues, proliferating cell nuclear antigen index of fibroblasts in granulation tissues, and the proliferating cell nuclear antigen index in the epidermis of HGF gene-transfer rats were significantly increased at 3 and 7 days after gene transfer. Semiquantitative reverse transcriptase-polymerase chain reaction revealed that the expression levels of transforming growth factor-beta1 and Colalpha2(I) mRNAs in the wounds of HGF gene-transfer rats were significantly decreased at 7 and 14 days, respectively. The hydroxyproline concentration in the wound was significantly less in HGF gene-transfer rats than in control rats at 3 days after gene transfer. These results suggest that HGF gene transfer into a skin wound may aid re-epithelialization and neovascularization in the early phase of wound healing, and that HGF may play a role in modulating cutaneous wound healing.

Defining a common region of DNA amplification at 22q11.2–12 in head and neck squamous cell carcinomas by quantitative FISH analysis

By comparative genomic hybridization (CGH) with 17 head and neck squamous cell carcinoma (HNSCC) cell lines, we previously detected an amplified region as a distinct peak at 22q11.2–12 in 3 cell lines. Because the possible presence of an oncogene was strongly suggested, the region was mapped in more detail by defining the minimum region that was commonly amplified by using fluorescence in situ hybridization (FISH) with a series of cosmids. Eighteen cosmid clones on 22q11.2–12 were assigned to their locations as a fractional length map and hybridized to cells from three HNSCC cell lines. The three cosmid clones, cHKA‐118, cHKAD‐26, and D22S938, showed the highest levels of amplification, and the size of the amplicon was calculated to be ∼ 1.7 Mb in the OM1 and HSC6 cell lines. Several genes related to oncogenesis, including PRKM1, map to this locus. Thus, the definition of the common region with the highest level of copy number increases by FISH provides a starting point for identifying the gene that may play an important role in the development of HNSCC.

Localization of carboxy-terminal type II procollagen peptide (pCOL-II-C) in diseased cartilage

A well characterized rabbit polyclonal antibody against human carboxy-terminal type II procollagen peptide (pCOL-II-C) was used to study the immunolocalization of pCOL-II-C in articular cartilage obtained from patients with osteoarthritis (OA), rheumatoid arthritis (RA), and control non-diseased joints. In moderately degenerative OA cartilage, immunoreactive chondrocytes were observed in all layers, particularly along the margins of fibrillation and fissures, in chondrocyte clusters and in osteochondrophytes. The grade of immunostaining in OA correlated directly with Mankins histological-histochemical scores of 0–7, but there was an inverse correlation between grade of immunostaining and Mankins scores of 8–14. The grade of immunostaining was significantly higher in OA than in RA and normal control cartilage. Since type II collagen is a unique component of articular cartilage, localization of pCOL-II-C could reflect the increased synthesis of type II collagen by chondrocytes in diseased cartilage.

Mechanism of metastasis of gastric cancer, from the viewpoint of basement membrane degradative enzyme.

上皮性癌の転移過程では, まず癌原発巣部における癌細胞の基底膜層の通過, 間質内での浸潤, その後に遭遇する脈管基底膜層の通過と脈管内への侵入, さらには転移としての着床部位における脈管外への遊出の際の脈管基底膜層の通過などが考えられ, 転移におけるいわゆるseed and soilの際に癌細胞が基底膜を何回にも渡って通過しなければならない.基底膜は主にIV型コラーゲンから構成されており, IV型コラーゲン分解酵素 (IVase) によって特異的に分解される.そこで癌組織中のIV ase活性はその癌の生物学的悪性度を表す指標となる可能性の追求を, まず組織中IV ase活性測定法を確立し, それを胃癌症例に用いて行い, その可能性を証明した.さらにIV型コラーゲンの分解産物である尿中3-hydroxyprolineが進行胃癌症例で増加していることもわかった.