Faculty Profiles - TAKIGAWA Masaharu

Papers - TAKIGAWA Masaharu

Division display 341 - 360 of about 473 ／ All the affair displays >>

軟骨由来の成長因子CTGF/Hcs24遺伝子に同定された新たな転写後制御エレメント,CAESAR

久保田聡, 近藤誠二, 江口傑徳, 服部高子, 中西徹, 滝川正春

日本骨代謝学会雑誌 18 ( 2 ) 119 - 119 2000.6

Expression of neurotrophins and their receptors (TRK) during fracture healing.

K Asaumi, T Nakanishi, H Asahara, H Inoue, M Takigawa

Bone 26 ( 6 ) 625 - 633 2000.6

Novel intracellular effects of human connective tissue growth factor expressed in Cos-7 cells

S Kubota, T Hattori, T Shimo, T Nakanishi, M Takigawa

FEBS LETTERS 474 ( 1 ) 58 - 62 2000.5

Language：English Publishing type：Research paper (scientific journal) Publisher：ELSEVIER SCIENCE BV

To clarify the multiple functionality of connective tissue growth factor (CTGF), we examined the effects of nascent CTGF within the cell by transient expression. Tn Cos-7 cells, expression of human CTGF induced an altered cell morphology, It was associated with an increased cellular DNA content and loose attachment, indicating the cells mere in G2/M phase. Overexpression of CTGF did not induce cell growth, whereas recombinant CTGF efficiently stimulated the proliferation extracellularly, These results indicate that intracellular CTGF may act as an antimitotic agent, thus it should also be noted that nascent CTGF was found to accumulate around the central mitotic machinery. (C) 2000 Federation of European Biochemical Societies.

Web of Science

researchmap

Ex vivo gene delivery using an adenovirus vector in treatment for cartilage defects

T Ikeda, T Kubo, T Nakanishi, Y Arai, K Kobayashi, O Mazda, S Ohashi, K Takahashi, J Imanishi, M Takigawa, Y Hirasawa

JOURNAL OF RHEUMATOLOGY 27 ( 4 ) 990 - 996 2000.4

Language：English Publishing type：Research paper (scientific journal) Publisher：J RHEUMATOL PUBL CO

Objective. To realize local selective gene expression in grafted chondrocytes for cartilage defect, we investigated the usefulness of an ex vivo gene delivery method using an adenovirus vector.
Methods. beta-galactosidase gene (LacZ) was transfected using an adenovirus vector to chondrocytes isolated from rat joints. The cells were then embedded into collagen gel, and LacZ expression in the gel was examined using 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside (X-gal) staining; beta-galactosidase activity was also measured. The collagen gel containing transfected chondrocytes was grafted to the experimental cartilage defects, and the expression of delivered gene was histologically examined after X-gal staining of the tissue containing the grafted area.
Results. X-gal positive chondrocytes in the gel accounted for 82% at one week and 55% at 8 weeks after gene delivery. beta-galactosidase activity decreased with time, but its expression was maintained even at 8 weeks after gene delivery. Chondrocytes used in the allograft maintained their morphology, and the expression of delivered gene continued during the 8 week period.
Conclusion. In this ex vivo method, delivered gene can be expressed efficiently for a long time; this method would be useful in allografts for cartilage defects.

Web of Science

researchmap

軟骨細胞の基質代謝に及ぼす周期的メカニカルストレスの影響

藤沢拓生, 服部高子, 窪木拓男, 矢谷博文, 山下敦, 滝川正春

日本顎関節学会雑誌 12 ( 1 ) 133 - 134 2000.4

Serum levels of connective tissue growth factor are elevated in patients with systemic sclerosis: Association with extent of skin sclerosis and severity of pulmonary fibrosis

S Sato, T Nagaoka, M Hasegawa, T Tamatani, T Nakanishi, M Takigawa, K Takehara

JOURNAL OF RHEUMATOLOGY 27 ( 1 ) 149 - 154 2000.1

Language：English Publishing type：Research paper (scientific journal) Publisher：J RHEUMATOL PUBL CO

Objective. To determine the serum levels and clinical correlation of connective tissue growth factors (CTGF) in patients with systemic sclerosis (SSc).
Methods. Serum samples from patients with limited cutaneous SSc (lSSc, n = 32), diffuse cutaneous SSc (dSSc, n = 28), systemic lupus erythematosus (SLE, n = 30), polymyositis/dermatomyositis (PM/DM, n = 20), and healthy control subjects (n = 30) were examined by ELISA for detection of CTGF.
Results. Serum CTGF levels in patients with SSc were significantly higher than those in patients with SLE or PM/DM, and in controls, CTGF levels in patients with dSSc were significantly higher than those in patients with lSSc. As for clinical correlation of CTGF, SSc patients with elevated CTGF had pulmonary fibrosis, decreased DLCO, and decreased vital capacity-more frequently than those with normal CTGF levels. Further, DLCO and vital capacity were inversely and directly correlated with serum CTGF levels in patients with SSc. The dSSc patients with disease duration of 1-3 years had significantly elevated levels of CTGF compared with dSSc patients with duration < 1 year or more than 3 years.
Conclusion. Serum CTGF levels were increased in patients with SSc, and correlated with the extent of skin sclerosis and the severity of pulmonary fibrosis. In addition, it appears that production of CTGF is involved in the development or maintenance of fibrosis rather than in initiation of fibrosis in SSc. These data suggest that CTGF plays a critical role in the development of fibrosis in SSc.

Web of Science

researchmap

Effects of CTGF/Hcs24, a product of a hypertrophic chondrocyte-specific gene, on the proliferation and differentiation of chondrocytes in culture

T Nakanishi, T Nishida, T Shimo, K Kobayashi, T Kubo, T Tamatani, K Tezuka, M Takigawa

ENDOCRINOLOGY 141 ( 1 ) 264 - 273 2000.1

Language：English Publishing type：Research paper (scientific journal) Publisher：ENDOCRINE SOC

Recently, we cloned a messenger RNA (mRNA) predominantly expressed in chondrocytes from a human chondrosarcoma-derived chondrocytic cell line, HCS-2/8, by differential display PCR and found that its gene, named hcs24, was identical with that of connective tissue growth factor (CTGF). Here we investigated CTGF/Hcs24 func- tion in the chondrocytic cell line HCS-2/8 and rabbit growth cartilage (RGC) cells. HCS-2/8 cells transfected with recombinant adenoviruses that generate CTGF/Hcs24 sense RNA (mRNA) proliferated more rapidly than HCS-2/8 cells transfected with control adenoviruses. HCS-2/8 cells transfected with recombinant adenoviruses that generate CTGF/Hcs24 sense RNA expressed more mRNA of aggrecan and type X collagen than the control cells. To elucidate the direct action of CTGF/Hcs24 on the cells, we transfected HeLa cells with CTGF/Hcs24 expression vectors, obtained stable transfectants, and purified recombinant CTGF/Hcs24 protein from conditioned medium of the transfectants. The recombinant CTGF/Hcs24 effectively promoted the proliferation of HCS-2/8 cells and RGC cells in a dose-dependent manner and also dose dependently increased proteoglycan synthesis in these cells. In addition, these stimulatory effects of CTGF/Hcs24 were neutralized by the addition of anti-CTGF antibodies. Furthermore, the recombinant CTGF/Hcs24 effectively increased alkaline phosphatase activity in RGC cells in culture. Moreover, RT-PCR analysis revealed that the recombinant CTGF/Hcs24 stimulated gene expression of aggrecan and collagen types II and X in RGC cells in culture. These results indicate that CTGF/Hcs24 directly promotes the proliferation and differentiation of chondrocytes.

Web of Science

researchmap

Rheumatoid arthritis-related antigen 47kDa (RA-A47) is a product of colligin-2 and acts as a human HSP47

T Hattori, K Takahashi, Y Yutani, T Fujisawa, T Nakanishi, M Takigawa

JOURNAL OF BONE AND MINERAL METABOLISM 18 ( 6 ) 328 - 334 2000

Language：English Publishing type：Research paper (scientific journal) Publisher：SPRINGER-VERLAG TOKYO

We previously isolated RA-A47, which is recognized as an antigen of rheumatoid arthritis (RA), from a human chondrosarcoma-derived cell line (HCS-2/8). The N-terminal 21-amino-acid sequence of RA-A47 had 81% homology to the deduced amino acid sequence of the human heat-shock protein (HSP) 47 gene, the colligin gene, and 100% homology to that of the colligin-2 gene. Moreover, as is HSP47, RA-A47 was a heat-inducible collagen-binding protein. To further characterize RA-A47, we isolated ra-a47 cDNA from HCS-2/8 cells and human periodontal ligament fibroblast (HPLF) cells. The isolated ra-a47 cDNAs from both cells were almost the same as that of collipin-2. C-504 and G(505) in the cDNA sequences of both cells and C-598 in the cDNA of HCS-2/8 were different from the corresponding bases of colligin-2 cDNA. These differences were also observed in genomic DNA. colligin cDNA was not isolated. To show that the isolated cDNA encodes RA-A47 protein, it was expressed in Cos-7 cells. The produced protein was 47 kDa and was recognized both with RA sera and antirat HSP47 antibody, indicating that it is RA-A47 and has structural similarity to HSP47. These results taken together with our previous finding show that RA-A47 is the putative colligin-2 gene product and behaves as a human HSP47. Although colligin has been considered the human HSP47 gene, failure to detect the colligin gene and its mRNA suggests that colligin does not exist in human cells and that the HSP47 gene is identical with colligin-2, which encodes RA-A47.

Web of Science

researchmap

Matrix Metalloproteinases and Tissue Inhibitors of Metalloproteinases in Synovial Fluids of Patients with Temporomandibular Joint Osteoarthritis

Manabu Kanyama, Takuo Kuboki, Shunji Kojima, Takuo Fujisawa, Takako Hattori, Masaharu Takigawa, Atsushi Yamashita

Journal of Orofacial Pain 14 ( 1 ) 20 - 30 2000

Language：English Publishing type：Research paper (scientific journal)

Aims: Imbalance between matrix metalloproteinases (MMPs) and their inhibitors (TIMPs) may be involved in the breakdown of articular cartilage matrix of the temporomandibular joint (TMJ). Aims: In this study, MMPs, TIMPs, and MMP-1/TIMP-1 complex levels were examined in TMJ synovial fluid samples aspirated from TMJ osteoarthritis (OA) patients (2 males, 8 females
mean age, 29.7 years) and asymptomatic control subjects (2 males, 8 females
mean age, 23.6 years) to determine the likelihood of increased proteolytic activity in the OA joints. Methods: The various types of MMPs and TIMPs were detected by Western blotting with monoclonal antibodies and gelatin zymography. The MMP-1/ TIMP-1 complex level was measured by an enzyme-linked immunosorbent assay kit. All aspirates were first analyzed for total protein content and then individually diluted to make the total protein levels equivalent. Results: The mean MMP-1/TIMP-1 complex concentration in the synovial fluids of the OA patients was 3.92 ± 1.39 ng/mL
this value was significantly lower (P &lt
0.05) than the value from control subjects (5.46 ± 1.32 ng/mL). Matrix metalloproteinase-1 (52 kDa), MMP-3 (57 kDa), TIMP-1 (28 kDa), and TIMP-2 (26 kDa) were detected in all of the normal and the OA samples. However, MMP-1 (28 kDa), MMP-2 (72 kDa), MMP-3 (45 kDa), and MMP-9 (83 kDa) were detected in higher concentration in the OA samples. Conclusion: These findings suggest a strong association between the OA-active joints and the presence of biologically active forms of known tissue degradation enzymes (MMP-1, MMP-3, and MMP-9).

Scopus

PubMed

researchmap

マトリクラインとMMP Invited

服部高子, 滝川正春

現代医療、現代医療社 (4), 909- 915, ( 4 ) 909 - 915 2000

Electrophoretic and serologic characterization of 56 kDa antigen (M56) with autologous serum derived from a chondrosarcoma patient: A shared antigen of immunoresponses in cancer and autoimmune diseases

K Fujiwara, H Udono, T Kunisada, A Kawai, H Inoue, M Takigawa, M Namba, E Nakayama

ELECTROPHORESIS 20 ( 17 ) 3335 - 3342 1999.11

Language：English Publishing type：Research paper (scientific journal) Publisher：WILEY-V C H VERLAG GMBH

We investigated whether antibodies specific to autologous cancer cells are produced in the peripheral blood of patients with chondrosarcoma. There have been few reports on the investigation of the immune responses, such as autologous antibody production, to chondrosarcoma. Here, tumor-associated antigens were separated by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis and detected by immunoenzymatic amplification. A 56 kDa molecule (M56) was detected in the serum from patients' peripheral blood. M56 is ubiquitously expressed in various kinds of tissue-derived cells. However, the molecule seemed to be retained mostly in the cytosolic compartment of lymphoid cells, while it was expressed on the cell surface of nonlymphoid cancer cells. Furthermore, the antibodies reactive to the 56 kDa molecule were frequently observed in sera derived from patients with other cancers and autoimmune diseases as compared to the sera from healthy control donors, suggesting that M56 is a common target molecule of immune responses in patients with various cancers and autoimmune diseases.

Web of Science

researchmap

Role and interaction of connective tissue growth factor with transforming growth factor-beta in persistent fibrosis: A mouse fibrosis model

T Mori, S Kawara, M Shinozaki, N Hayashi, T Kakinuma, A Igarashi, M Takigawa, T Nakanishi, K Takehara

JOURNAL OF CELLULAR PHYSIOLOGY 181 ( 1 ) 153 - 159 1999.10

Language：English Publishing type：Research paper (scientific journal) Publisher：WILEY-LISS

Skin fibrotic disorders are understood to develop under the influence of some growth factors, such as transforming growth factor-beta (TCF-beta), basic fibroblast growth factor (bFGF), or connective tissue growth factor (CTGF). To establish an appropriate animal model of skin fibrosis by exogenous application of growth factors, we investigated the in vivo effects of growth factors by injecting TCF-beta, CTGF, and bFGF into the subcutaneous tissue of newborn mice. A single application of TGF-beta or bFGF resulted in the formation of transient granulated tissue that disappeared despite 7 days of consecutive injections. A single CTGF injection also caused slight granulation. However, injecting TGF-beta plus CTGF produced long-term fibrotic tissue, which persisted for at least 14 days. Also, fibrotic tissue was observed when CTGF was injected from 4 to 7 days after TGF-beta injections for the first 1-3 days. In situ hybridization analysis revealed the expression of CTGF mRNA in the fibroblasts at least in a few fibrotic conditions. These findings suggest that either CTGF mRNA or an application of exogenous CTGF protein is required for the development of persistent fibrosis. From our study, it appears that interaction of several growth factors is required for persistent fibrotic tissue formation, with TGF-beta causing the induction and CTGF needed for maintenance of skin fibrosis. The animal model on skin fibrosis by exogenous application of growth factors developed in this study may prove useful for future studies on fibrotic disorders. J. Cell. Physiol. 181:153-159, 1999. (C) 1999 Wiley-Liss, Inc.

Web of Science

researchmap

Direct adenovirus-mediated gene delivery to the temporomandibular joint in guinea-pigs

T Kuboki, T Nakanishi, M Kanyama, W Sonoyama, T Fujisawa, K Kobayashi, T Ikeda, T Kubo, A Yamashita, M Takigawa

ARCHIVES OF ORAL BIOLOGY 44 ( 9 ) 701 - 709 1999.9

Language：English Publishing type：Research paper (scientific journal) Publisher：PERGAMON-ELSEVIER SCIENCE LTD

Adenovirus vector system is expected to be useful for direct gene therapy for joint disease. This study first sought to confirm that foreign genes can be transferred to articular chondrocytes in primary culture. Next, recombinant adenovirus vectors harbouring beta-galactosidase gene (LacZ) was injected directly into the temporomandibular joints of Hartley guinea-pigs to clarify the in vivo transfer availability of the adenovirus vectors. Specifically, recombinant adenovirus harbouring LacZ gene (AxlCALacZ) was injected into the upper joint cavities of both mandibular joints of four male 6-week-old Hartley guinea-pigs. Either the same amount of recombinant adenovirus without LacZ gene (Axlw) suspension (placebo) or the same amount of phosphate-buffered saline solution (control) were injected into the upper joint cavities of both joints of another four male guinea-pigs. At 1, 2, 3 and 4 weeks after injection, the joints were dissected and the expression of delivered LacZ was examined by 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside (X-gal) staining and reverse transcriptase-polymerase chain reaction (RT-PCR). To investigate the expression of transferred gene in other organs, total RNA was extracted from liver, kidney, heart and brain and the expression of LacZ mRNA and 18 S ribosomal RNA were analysed by RT-PCR. Clear expression of LacZ was observed in the articular surfaces of the temporal tubercle, articular disc and synovium of the temporomandibular joints even 4 weeks after injection in the AxlCALacZ-injected group, while no expression was detected in placebo and control groups. Histological examination confirmed that LacZ activity was clearly detected in a few cell layers of the articular surface tissues, which is much more efficient than in a previously study of the knee joint. In the other organs, expression of the delivered transgene was not observed. Based on these findings, direct gene delivery into the articular surface of the temporomandibular joint using the adenovirus vector is feasible as an effective in vivo method, (C) 1999 Elsevier Science Ltd. All rights reserved.

Web of Science

researchmap

Control of delivered gene expression in chondrocytes using heat shock protein 70B promoter

Y Arai, T Kubo, K Kobayashi, T Ikeda, K Takahashi, M Takigawa, J Imanishi, Y Hirasawa

JOURNAL OF RHEUMATOLOGY 26 ( 8 ) 1769 - 1774 1999.8

Language：English Publishing type：Research paper (scientific journal) Publisher：J RHEUMATOL PUBL CO

Objective. To investigate whether the expressions of delivered Escherichia coli beta-galactosidase (LacZ) gene and transforming growth factor-beta 1 (TGF-beta 1) gene are regulated by the stress response of human chondrocyte-like cells (HCS-2/8) when heat shock protein 70B (HSP70B) promoter is inserted into the adenovirus vector,
Methods. Two adenovirus vectors that contain either LacZ gene or TGF-beta 1 gene regulated by HSP70B promoter were constructed. One of the adenovirus vectors was added to the culture of HCS-2/8 and gene transduced cells were produced. We applied heat stress (43 degrees C) to the transduced cells for 2 h and examined whether the expression of transduced LacZ and TGF-beta 1 genes is affected by the stress, using 5 bromo-4-chloro-3-indolyl-beta-D-galactopyranoside (X-gal) staining, measurement of B-galactosidase activity, Northern blotting, and ELISA.
Results. The percentage of X-gal positive stained cells in LacZ gene-delivered cells with heat stress was significantly higher than in controls (no heat stress). With heat stress, beta-galactosidase activity increased significantly, and the band of exogenous TGF-beta 1 mRNA became more apparent and the expression was maintained during the 24 h monitoring period. TGF-beta 1 level in culture supernatant of TGF-beta 1 gene-delivered cells with heat stress (5377.3 +/- 321.1 pg/ml) was significantly higher than in the controls (853.2 +/- 29.2 pg/ml).
Conclusion, HSP70B promoter could regulate the expression of delivered genes according to the intensity of heat stress.

Web of Science

researchmap

骨・軟骨の発生過程における軟骨由来成長因子CTGF/Hcs24の発現とCbfa1によるその制御

中西徹, 浅野将宏, 縄稚久美子, 山合友一郎, 小守寿文, 西田崇, 吉道玄, 久保田聡, 服部高子, 滝川正春

生化学 71 ( 8 ) 1033 - 1033 1999.8

TNFαによるRA-A47の軟骨細胞内局在の変化と抗原提示

服部高子, 藤沢拓生, 油谷安孝, 中西徹, 滝川正春

生化学 71 ( 8 ) 969 - 969 1999.8

Immunohistochemical localization of connective tissue growth factor in the rat central nervous system

Y Kondo, T Nakanishi, M Takigawa, N Ogawa

BRAIN RESEARCH 834 ( 1-2 ) 146 - 151 1999.7

Language：English Publishing type：Research paper (scientific journal) Publisher：ELSEVIER SCIENCE BV

Connective tissue growth factor (CTGF) is an immediate early growth-responsive gene but its distribution and significance in the central nervous system (CNS) are unknown. We investigated the distribution of CTGF-like immunoreactivity (CTGF-IR) in the rat CNS using a specific antiserum against CTGF oligopeptide. The majority of CTGF-IR was observed in astrocytes. Ependymal cells lining the wall of the cerebral ventricle and tanycytes Lining the central canal of the spinal cord showed the strongest CTGF-IR, while there was a diffuse but weak signal in the gray matter of the spinal cord. CTGF-IR was also detected in the cytoplasm of a subpopulation of pyramidal neurons in the cerebral cortex. Our results showed that CTGF-IR is widely distributed in the CNS at bath regional and cellular levels, suggesting a complex functional role in the CNS. (C) 1999 Elsevier Science B.V. All rights reserved.

Web of Science

researchmap

Connective tissue growth factor induces the proliferation, migration, and tube formation of vascular endothelial cells in vitro, and angiogenesis in vivo

T Shimo, T Nakanishi, T Nishida, R Asano, M Kanyama, T Kuboki, T Tamatani, K Tezuka, M Takemura, T Matsumura, M Takigawa

JOURNAL OF BIOCHEMISTRY 126 ( 1 ) 137 - 145 1999.7

Language：English Publishing type：Research paper (scientific journal) Publisher：JAPANESE BIOCHEMICAL SOC

Connective tissue growth factor (CTGF) is a novel cysteine-rich, secreted protein. Recently, we found that inhibition of the endogenous expression of CTGF by its antisense oligonucleotide and antisense RNA suppresses the proliferation and migration of vascular endothelial cells. In the present study, the following observations demonstrated the angiogenic function of CTGF in vitro and in vivo: (i) purified recombinant CTGF (rCTGF) promoted the adhesion, proliferation and migration of vascular endothelial cells in a dose-dependent manner under serum-free conditions, and these effects were inhibited by anti-CTGF antibodies; (ii) rCTGF markedly induced the tube formation of vascular endothelial cells, and this effect was stronger than that of basic fibroblast growth factor or vascular endothelial growth factor; (iii) application of rCTGF to the chicken chorioallantoic membrane resulted in a gross angiogenic response, and this effect was also inhibited by anti-CTGF antibodies, (iv) rCTGF injected with collagen gel into the backs of mice induced strong angiogenesis in vivo. These findings indicate that CTGF is a novel, potent angiogenesis factor which functions in multi-stages in this process.

Web of Science

researchmap

Physiological function of connective tissue growth factor (CTGF/Hcs24) - Its roles in the process of endochondral ossification Reviewed

T Nakanishi, M Takigawa

SEIKAGAKU 71 ( 6 ) 429 - 432 1999.6

骨芽細胞様細胞株MC3T3-E1のメカニカルストレスに対する応答性神経栄養因子の役割

稲熊尚広, 山本照子, 中西徹, 山城隆, 山下和夫, 滝川正春

日本骨代謝学会雑誌 17 ( 2 ) 195 - 195 1999.6

　 PREV - NEXT 　

< TAKIGAWA Masaharu >

Papers - TAKIGAWA Masaharu