Atsushi Jikuhara

MAP kinase-mediated proliferation of DLD-1 carcinoma by the stimulation of protease-activated receptor 2.

Protease-activated receptor-2 (PAR-2) has been demonstrated to be highly expressed in the gastrointestinal tract. In the present study, we investigated the effects of PAR-2 stimulation on the cell signaling and proliferation of DLD-1, a human colon carcinoma cell line, in comparison with the PAR-1 stimulation. PAR-2 stimulation by agonist peptide SLIGKV concentration-dependently induced the increase in [Ca2+]i and the proliferation of DLD-1 whereas the inverse peptide LSIGKV did not. Trypin (10(-9) M), an agonist protease for PAR-2, also enhanced the proliferation of DLD-1. The proliferative response of DLD-1 to PAR-2 stimulation was associated with the transient phosphorylation of MEK and MAP kinase, but not p38 MAP kinase and JNK. Inhibition of MEK by PD98059 (50 microM) completely inhibited the proliferation-stimulating effects as well as the phosphorylation of MAP kinase induced by PAR-2 agonist peptide (100 microM) and trypsin (10(-9) M). The prolonged treatment with PAR-2 agonist peptide for more than one hour was required for the enhanced proliferative response, suggesting the existence of unknown long-lasting cooperative signaling with MAP kinase cascade. PAR-1 stimulation by the agonist peptide SFLLRN (100 microM) or thrombin (10(-8) M) produced Ca2+ signaling, however, the stimulation neither produced the cell proliferative response nor the activation of MEK-MAP kinase cascade. These results indicated that Ca2+ signaling induced by PARs activation was not enough for inducing the cell proliferation in DLD-1 cells and that stimulation of PAR-2 can induce the activation of MEK-MAP kinase cascade, leading to the growth promoting response.

The effects of stimulating protease-activated receptor-1 and -2 in A172 human glioblastoma.

SummaryHuman glioblastoma cell line A172 expressed protease-activated receptor-1 and -2 (PAR-1 and PAR-2). We investigated the effects of the stimulation of these receptors by receptor-activating agonist peptides on the Ca2+ signaling, protein kinase C translocation, cell morphology and cell proliferation in A172. Both PAR-1 agonist SFLLRN and PAR-2 agonist SLIGKV induced an increase in [Ca2+]i. The prior treatment of A172 with PAR-2 agonist SLIGKV did not influence the [Ca2+]i response to PAR-1 agonist SFLLRN or thrombin, however, the prior treatment with PAR-1 agonist SFLLRN or thrombin completely abolished the second response to PAR-2 agonist SLIGKV. Treatment with each agonist peptide produced thinner and fewer processes in A172. The PAR-2 agonist inhibited the proliferation of A172 significantly while PAR-1 agonist did not. PKC-α and γ were translocated from cytosol to membrane with either PAR-1 or PAR-2 stimulation, however, ı was specifically translocated with SFLLRN, and λ with SLIGKV, respectively. These results indicated that PAR-1 and PAR-2 stimulation produced a similar [Ca2+]i response and morphological changes in A172 glioblastoma while the effects on the cell proliferation and activation of PKC isozymes were distinct, suggesting that different signal transduction pathways were activated by these receptors. The uni-directional cross desensitization implies a functional linkage between PAR-1 and PAR-2 receptors.

Immunohistochemical studies of colorectal cancers that developed following pelvic irradiation.

Radiation therapy is believed to be a predisposing factor in the development of carcinoma. During the period from 1974 to 1997, we treated five colorectal cancers in four patients who had undergone pelvic irradiation for cervical cancer. Radiation-induced changes in the remaining bowel were recognized in all of these patients. Of the five tumors, two were histologically diagnosed as mucinous adenocarcinoma, demonstrating a higher distribution than usual: To clarify the characteristics of the tumors, we examined the frequency of mutation in K-ras and p53 genes, and the proliferating cell nuclear antigen (PCNA) proliferation activity. The K-ras and PCNA stains did not show any clear characteristics of radiation-induced colorectal cancers in comparison with ordinary colorectal cancers; however, it was of interest that strongly positive staining was observed in our mucinous adenocarcinomas, even though a low frequency of p53 gene mutation has been reported in ordinary mucinous adenocarcinomas.

Crohn’s disease mimicking as bowel endometriosis

Abstract We report a 27-year-old female with Crohn’s disease clinically misdiagnosed with intestinal endometriosis. Her complaints were abdominal pain and fullness, which occurred monthly during her menstual period. Although we had no histopathological evidence, we diagnosed her as bowel endometriosis on the basis of her clinical course. Since nafarelin acetate therapy started, the symptoms due to mechanical subileus have improved. The transverse colon, a 70 cm segment of the ileum, including the terminal ileum, were resected because of repeated symptoms of bowel obstruction despite prolonged nafarelin therapy. Histopathological findings of the resected specimen revealed Crohn’s disease without endometrial tissue. In our patient, an increased cortisol and ACTH secretion, a side effect of nafarelin, was noted during the therapy. This case showed that nafarelin therapy could increase serum concentration of ACTH and cortisol, which was considered to suppress the pathology of Crohn’s disease by its anti-inflammatory action. We emphasize that intestinal examination must be performed with Crohn’s disease in mind, even if nafarelin acetate is effective.