Kazuo Ohmori

The Association of Lumbar Disc Disease with Vitamin-D Receptor Gene Polymorphism

Background: Although the etiology of lumbar disc disease is unknown, it has been suggested that a genetic factor contributes to its development. Recently, some genetic polymorphisms have been found to be related to clinical disorders. We investigated the association between vitamin-D receptor gene and estrogen receptor gene polymorphisms and lumbar disc disease in young adults.Methods: The participants included 205 young adults (166 women and thirty-nine men) with or without low-back problems. A magnetic resonance imaging scan of the lumbar spine was performed for all subjects, and the grade of disc degeneration was determined, according to the four-grade classification system of Schneiderman et al. The presence or absence of disc herniation was also evaluated. Genomic DNA was extracted from peripheral blood samples. The polymorphisms of the vitamin-D receptor and estrogen receptor genes were detected with use of a polymerase-chain-reaction assay. The restriction fragment length polymorphisms (RFLPs) for the vitamin-D receptor gene were analyzed by TaqI and ApaI restriction enzymes. XbaI and PvuII restriction enzymes were used for the estrogen receptor gene analysis. The distribution of polymorphism in subjects with disc degeneration and/or disc herniation was compared with that in the normal subjects.Results: The allelic frequencies of both vitamin-D receptor gene and estrogen receptor gene polymorphisms were similar to those in previous analyses of Japanese subjects. The allelic variation in the vitamin-D receptor gene was associated with multilevel and severe disc degeneration and disc herniation. The Tt allele was found to be more frequently associated with multilevel disc disease, severe disc degeneration, and disc herniation than was the TT allele. No additional associations were found.Conclusions: This study revealed that the Tt allele of the vitamin-D receptor gene was more frequently associated with multilevel and severe disc degeneration and disc herniation than was the TT allele, pointing to an increased risk of disc disease at an early age in subjects with the Tt allele in the vitamin-D receptor gene.

Pathomechanism of Myelopathy and Surgical Results of Laminoplasty in Elderly Patients With Cervical Spondylosis

Study Design. Clinical and radiologic analyses in elderly patients with cervical myelopathy. Objective. To investigate the pathomechanism of cervical myelopathy and the surgical results of laminoplasty in elderly patients older than 70 years old. Summary of Background Data. To date, there have been no reports on the pathomechanism of cervical myelopathy in elderly patients. Further, the surgical results and postoperative complications of laminoplasty have not been fully evaluated in elderly patients. Methods. Eighty-nine patients who underwent cervical laminoplasty were reviewed. The patients were divided into 2 groups according to the age at the time of operation (the elderly patient group: 20 patients who were older than 70 years old, and the control group: 69 patients who were younger than 69 years old). Pre- and postoperative neurologic status (the Japanese Orthopedic Association score) and postoperative complications were compared between the two groups. Radiologic features were also examined. Results. There was no statistical difference in the recovery rate of Japanese Orthopedic Association score between the elderly patient group and the control group. Activities of daily living improved in the elderly patients. Several complications, such as delirium and worsening hypertension, were found in the elderly patient group. In the preoperative radiographs, the incidence of either retrolisthesis or anterolisthesis in the elderly patient group was significantly higher than that in the control group. Conclusions. Retrolisthesis and anterolisthesis are often the cause of myelopathy in elderly patients. Surgical decompression for cervical myelopathy was beneficial even in elderly patients older than 70 years old. Laminoplasty achieves stability of the cervical spine, and this procedure is reasonable for the treatment.

Effects of Epidermal Growth Factor on Invasiveness through the Extracellular Matrix in High‐ and Low‐metastatic Clones of RCT Sarcoma in vitro

We investigated the invasiveness of tumor cells through the extracellular matrix and the influence of epidermal growth factor (EGF) on tumor cell invasion using in vitro systems in high‐[RCT(+)] and low‐metastatic [RCT(–)] clones established from poorly differentiated murine RCT sarcoma in C3H/He mice. In the invasion assay using a filter coated with reconstituted basement membrane (Matrigel) in a Boyden chamber, RCT(+) cells were more invasive than RCT(‐) cells. The attachment of RCT(+) cells to extracellular matrix components and the degradation of type IV collagen by the cells were significantly greater than with RCT(–) cells. However, there was no significant difference in the migration of cells to the extracellular matrix components between cultured RCT(+) and RCT(‐) cells. These findings suggested that the different invasiveness of these clone cells was associated with the difference in the ability of attachment to and degradation of the matrix. The level of laminin receptor expression in RCT(+) cells was about four‐fold that in RCT(–) cells and laminin stimulated the type IV collagenolytic activity of RCT(+) cells, suggesting that RCT(+) cell attachment to laminin via laminin receptor on the cell surface induced the production of type IV collagenase by the tumor cells. EGF did not affect the invasiveness of RCT(–) cells. In RCT(+) cells, EGF stimulated the invasiveness through Matrigel, the attachment to extracellular matrix components and the degradation of type IV collagen through high‐affinity EGF receptors (EGFR), with Kt of pM order, while the migration to the matrix was not influenced by EGF. These findings suggest that the stimulatory effect of EGF on invasion is related to the acceleration of cell adhesion, and the degradative cascade of the extracellular matrix and high‐affinity EGFRs play an important role in the effect of EGF on in vitro invasiveness in this tumor.

Cystic spinal cord tumors : Magnetic resonance imaging correlated to histopathological findings

We report the characteristics of magnetic resonance imaging (MRI) of cystic intradural extramedullary spinal cord tumors (cystic neurilemmoma, epidermoid cyst, and enterogeneous cyst). T1-weighted MRI enhanced with gadolinium-DTPA clearly demonstrated the rim morphology of these tumors. The comparison between the rim enhancement pattern and histopathological findings offered possible qualitative diagnosis of these cystic spinal cord tumors by MRI.

Tumor cell attachment to laminin promotes degradation of the extracellular matrix and cell migration in high-metastatic clone cells of RCT sarcoma in vitro.

We investigated the roles of extracellular matrix proteins, iaminin and fibronectin, in promoting invasiveness through the extracellular matrix in high‐metastatic [RCT(+)] clone cells established from poorly differentiated murine RCT sarcoma in C3H/He mice. Laminin stimulated the type IV collagenolytic activity of RCT(+) cells. After more than 6 h of incubation, the type IV collagenolysis of the cell‐conditioned medium was significantly higher in laminin‐treated groups compared with the control. The migration activity of RCT(+) cells was stimulated by laminin. However, fibronectin did not influence the type IV collagenolysis or cell migration in this clone cell. The amino acid sequence YIGSR, which is derived from laminin, inhibited the laminin‐mediated cell attachment and the laminin‐promoted type IV collagenolysis, as well as cell migration of RCT(+) cells. RGD derived from fibronectin did not influence the cell attachment to laminin or Matrigel in this clone. In the invasion assay employing a Matrigel coated filter in a Boyden chamber, YIGSR showed greater inhibition of invasion through the Matrigel than did RGD with RCT(+) cells. YIGSR might inhibit the promoted‐matrix degradation and cell migration in response to the cell attachment to laminin by competing with laminin for binding to cell surface laminin receptor. We suggest that laminin‐mediated cell attachment to the extracellular matrix may play a role in promoting the matrix degradation and cell migration during metastatic cascades.

Prognostic Value of the Doubling Time of Serum C‐reactive Protein and Alkaline Phosphatase Levels in Primary Bone and Soft Tissue Tumors

We investigated the clinical relevance of doubling time (DT) of serum laboratory data obtained in routine clinical examination of patients with primary bone and soft tissue tumors, in comparison with major clinical and pathological parameters (age at presentation, sex, tumor size, location, clinical stage and histologic grade) by uni‐ and multivariate analyses. In 64 patients with primary bone and soft tissue tumors (primary bone tumors: 39, primary soft tissue tumors: 25) and 68 cancer patients, the pretreatment DT values of serum C‐reactive protein (CRP), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), calcium (Ca), phosphate (P) levels were measured, as well as the erythrocyte sedimentation rate (ESR: mm/60 min); these values were then compared with overall survival, local recurrence‐free survival and metastasis‐free survival. Only DT of CRP and ALP (CRP‐DT, ALP‐DT) were found to be correlated with disease outcome in patients with primary bone and soft tissue tumors. In cancer patients, only CRP‐DT showed a relation with clinical stage and histologic grade, but the ALP‐DT in patients with bone metastasis was significantly shorter than that in patients with metastases at other sites or in those with no metastasis. Among all tumor patients, those with bone metastasis showed the shortest ALP‐DT compared with those with lung, liver and brain metastasis. Univariate analysis showed that shorter CRP‐DT and ALP‐DT are associated with poor overall survival, and the development of local recurrence and metastasis. These findings suggest that pretreatment CRP‐ and ALP‐DT could be additional prognostic parameters for disease outcome in patients with primary malignant bone and soft tissue tumors. However, in multivariate analysis, only ALP‐DT, but not CRP‐DT, was an independent prognostic parameter for these disease outcomes.

Regulation of Gelatinase Production and Invasiveness by Organ-Specific Fibroblasts in High- and Low-Metastatic Clones from Murine RCT Sarcoma

Regulation of gelatinase production, invasiveness and migration activity by organ-specific fibroblasts from embryo, subcutaneous and lung tissues of mice were investigated in high-metastatic RCT+ and low-metastatic RCT- clones established from a poorly differentiated murine sarcoma. In the conditioned media of RCT+ cells, mouse skin fibroblasts (MSF) obtained from the tissue of tumor origin (orthotopic) stimulated the production of the 105-kD gelatinase more than C3H/ 10T1/2 clone 8 (C3H/10 T1/2 CL8) or mouse lung fibroblasts (MLF). In the conditioned media of RCT- cells, however, cocultivation with fibroblasts showed only slight stimulatory effects on the production of the 105-kD gelatinase. In the invasion assay, using a reconstituted basement membrane (matrigel), RCT+ cells cocultivated with MSF showed significantly higher invasiveness than those cocultivated with C3H/10T1/2 CL8 or MLF. However, no significant differences were shown in the invasiveness of RCT- cells in cocultivation with three types of fibroblasts and in cultivation without fibroblasts. There was no significant difference in migration activity between RCT+ and RCT- cells cultivated alone. But in the cocultivation of both clones with MSF, the migration activity of RCT+ cells was significantly higher than that of RCT- cells. These findings suggest that MSF might delineate the difference in characteristics related to the metastatic potential of RCT+ and RCT- cells through regulation by organ-specific factors.

Characteristics of high and low laminin-adherent Dunn osteosarcoma cells selected by adhesiveness to laminin. Correlation between invasiveness through the extracellular matrix and pulmonary metastatic potential.

We investigated the invasiveness through the extracellular matrix and pulmonary metastatic potential in high laminin-adherent [LN(+)] and low laminin-adherent [LN(-)] Dunn oseosarcoma cells selected for adhesiveness to laminin. In the invasion assay using a reconstituted basement membrane (matrigel) in a Boyden chamber, LN(+) cells proved to be more invasive than LN(-) and the parental Dunn cells. Pulmonary metastatic potential was correlated with invasiveness through the matrigel in three cell types. The ability of LN(+) cells to attach to laminin and the matrigel was significantly higher than that of LN(-) or the parental Dunn cells. LN(-) cells showed much lower attachment ability compared to the other cells. There were no significant differences in type IV collagenolysis and cell migration among the three cell types. In LN(+) and the parental Dunn cells, laminin significantly stimulated type IV collagenolytic and migration activities. LN(-) cells showed no significant differences of type IV collagenolysis and cell migration in response to laminin. These findings suggested that the different invasiveness of these cells was associated with the difference in the abilities of cell attachment to laminin and type IV collagenolysis and cell migration activities stimulated by laminin. YIGSR inhibited not only the laminin-mediated cell attachment but also the type IV collagenolysis and cell migration induced by the cell attachment to laminin. The amount of laminin receptor of LN(+) cells was about fourfold that of LN(-) and twofold that of the parental Dunn cells, suggesting the stimulatory effect of laminin on the invasiveness through the matrix is associated with the level of laminin receptor expression in these cells. The present study provides several findings suggesting that laminin has important roles in invasiveness through the extracellular matrix and metastasis formation in Dunn osteosarcoma cells.