Ferry Sandra

RANKL-induced DC-STAMP is essential for osteoclastogenesis.

Osteoclasts are bone-resorbing, multinucleated giant cells that are essential for bone remodeling and are formed through cell fusion of mononuclear precursor cells. Although receptor activator of nuclear factor–κB ligand (RANKL) has been demonstrated to be an important osteoclastogenic cytokine, the cell surface molecules involved in osteoclastogenesis are mostly unknown. Here, we report that the seven-transmembrane receptor-like molecule, dendritic cell–specific transmembrane protein (DC-STAMP) is involved in osteoclastogenesis. Expression of DC-STAMP is rapidly induced in osteoclast precursor cells by RANKL and other osteoclastogenic stimulations. Targeted inhibition of DC-STAMP by small interfering RNAs and specific antibody markedly suppressed the formation of multinucleated osteoclast-like cells. Overexpression of DC-STAMP enhanced osteoclastogenesis in the presence of RANKL. Furthermore, DC-STAMP directly induced the expression of the osteoclast marker tartrate-resistant acid phosphatase. These data demonstrate for the first time that DC-STAMP has an essential role in osteoclastogenesis.

Immunohistochemical evaluation of PCNA and Ki-67 in ameloblastoma

Thirty-two ameloblastoma tissues were immunohistochemically studied using monoclonal anti-proliferating cell nuclear antigen (PCNA) and anti-Ki-67 antibodies. Positive cells were evaluated and analyzed in relation to the WHO classification, cytological pattern of the outer layer cell, clinical appearance, tumor location, radiographic appearance and patients age. In regard to the cytological pattern of the outer layer cells, the basal cell type had significantly higher PCNA and Ki-67 (P<0.05) labeling indices than the cuboidal cell type. The solid type had significantly higher PCNA and Ki-67 (P<0.05) labeling indices than the cystic and the mixed type. The labeling index of the younger patient was found to be the lowest, the middle age was in the middle and the older patient was the highest. These results indicated that the proliferating activities of ameloblastomas are quite variable, and the evaluations of Ki-67 and PCNA seem to be good indicators to assess the proliferating activity of each type of ameloblastomas.

Two relatively distinct patterns of ameloblastoma: an anti‐apoptotic proliferating site in the outer layer (periphery) and a pro‐apoptotic differentiating site in the inner layer (centre)

Two relatively distinct patterns of ameloblastoma: an anti‐apoptotic proliferating site in the outer layer (periphery) and a pro‐apoptotic differentiating site in the inner layer (centre)

Possible involvement of MIP-1α in the recruitment of osteoclast progenitors to the distal tibia in rats with adjuvant-induced arthritis

In the rat model of rheumatoid arthritis, a marked formation of osteoclasts is found in the distal tibia and the metatarsal bone. It was therefore postulated that osteoclast progenitors would be increased in the bone marrow cavities of rats with adjuvant-induced arthritis (AA rats). Bone marrow cells obtained from tibia of AA rats were cultured to form cells in the osteoclast lineage to access the number of osteoclast progenitors. Unexpectedly, only a suppressed level of osteoclast progenitors was detected in the diaphyseal bone marrow of tibia in AA rats. Distribution of osteoclast progenitors in the bone marrow cavity was examined, and it was shown that osteoclast progenitors accumulated in the distal tibia. Macrophage inflammatory protein (MIP)-1α, an osteoclastogenic CC chemokine, was expressed in ED-1-positive macrophages localizing in the distal tibia with marked bone destruction. Chemotaxis studies showed that MIP-1α expressed significant activity towards bone marrow cells. The suppressed level of osteoclastogenesis in bone marrow cells of AA rats was restored to a normal level by the addition of MIP-1α. It was suggested that MIP-1α is involved in the migration of osteoclast progenitors to the distal tibia as well as in osteoclastogenesis in AA rats. In these rats, in situ hybridization of the distal tibia with a high level of bone destruction showed significant expression of Receptor activator nuclear factor κB ligand (RANKL) messenger RNA in aggregates of multinucleated osteoclast-like cells present in the bone marrow cavity, a unique pathological feature for these rats. Migrated osteoclast progenitors are thought to be efficiently differentiated into osteoclasts in response to RANKL expressed by the aggregates of osteoclast-like cells under the influence of the MIP-1α. Such positive-feedback regulation of osteoclastogenesis could result in the highest recruitment of active osteoclasts in the area of marked bone destruction.

TNF inhibited the apoptosis by activation of Akt serine/threonine kinase in the human head and neck squamous cell carcinoma

Tumour necrosis factor (TNF) is known to induce apoptosis, but recently, TNF was shown to promote cell survival, a process regulated by phosphatidylinositol-3-OH kinase (PI3K) and the NFkappaB pathway. In this study, we investigated the relationship between the molecules implicated in regulating TNF-induced cell survival and apoptosis induced by TNF in a human head and neck squamous cell carcinoma cell line (SAS), with special reference to the Akt pathway, one of the pathways related to cell survival. In SAS cells, TNF induced the phosphorylation of Akt at both Ser473 and Thr308, causing the activation of Akt, and also induced the phosphorylation and degradation of IkappaB (inhibitor of NFkappaB). This phosphorylation and degradation was inhibited by pretreating the cells with the PI3K inhibitors, wortmannin or LY294002. The apoptosis of SAS cells induced by TNF was dependent on the concentration: a high concentration of TNF, but not a low concentration, induced apoptosis within 30 h. However, a low concentration of TNF in the presence of wortmannin or LY294002 induced apoptosis. Furthermore, expression of the kinase-negative form of Akt, IKKalpha or IKKbeta, and the undegradable mutant of IkappaB, also induced apoptosis at low concentrations of TNF. When the SAS cells expressed constitutively activated Akt, apoptosis was not induced, even by high concentrations of TNF. These observations suggest that, in the SAS cell line, the PI3K-NFkappaB pathway contributes to TNF-induced cell survival and that inhibition of this pathway accelerates apoptosis.

The role of MDM2 in the proliferative activity of ameloblastoma.

Ameloblastoma is a unique tumor in the oral and maxillofacial region with various levels of proliferative activity in each type. p53 is most commonly found to be mutated in human cancer and sometimes is overexpressed also in other lesions, such as ameloblastoma. Murine Double Minute 2 (MDM2) is able to physically associate with the p53 tumor suppressor and therefore block the growth suppressive functions of p53. In the present study, immunohistochemistry, western blotting and enzyme-linked immunosorbent assay for p53 mutant selective test were done. MDM2 was overexpressed in ameloblastoma and the results showed different numbers of MDM2 labeling index based on both WHO classification and cytological pattern of outer layer cells. Basal ameloblastoma, which has a high proliferative activity, had the highest MDM2 labeling index. We suggest MDM2 protein caused the high proliferative activity of ameloblastoma, especially in basal cell ameloblastoma.

Regulation of osteoclastogenesis by simon extracts composed of caffeic acid and related compounds : successful suppression of bone destruction accompanied with adjuvant-induced arthritis in rats

Simon extracts are vitamin K1-rich food materials extracted from the leaves of the Simon sweet potato. Although vitamin K is known to stimulate bone formation, we postulated that Simon extracts also contain unknown biological compounds having the ability to regulate bone resorption. Here we prepared the vitamin K-free fraction from the Simon extracts and investigated the ability of this fraction on the differentiation of osteoclasts. A remarkable inhibitory effect of osteoclastogenesis was observed when osteoclast precursors were treated with this fraction in rat bone marrow culture systems as well as in a pure differentiation system using murine osteoclast precursor cell line. The vitamin K-free Simon extracts markedly suppressed severe bone destruction mediated by abundant osteoclasts associated with adjuvant-induced arthritis in rats. High performance liquid chromatography (HPLC) analysis revealed that the vitamin K-free Simon extracts contained three types of low molecular weight inhibitors for osteoclastogenesis; caffeic acid, chlorogenic acids and isochlorogenic acids. Among these substances, caffeic acid showed the most powerful inhibitory effects on osteoclastogenesis. Caffeic acid significantly suppressed expression of NFATc1, a key transcription factor for the induction of osteoclastogenesis. Our current study enlightened a high utility of the Simon extracts and their chemical components as effective regulators for bone resorption accompanied with inflammation and metabolic bone diseases.

TRAIL Cleaves Caspase-8, -9 and -3 of AM-1 Cells: A Possible Pathway for TRAIL to Induce Apoptosis in Ameloblastoma

Tumor-necrosis-factor-related apoptosis-inducing ligand (TRAIL/Apo-2L), a potent ligand in inducing apoptosis, has recently emerged as a novel anticancer agent based on its ability to induce apoptosis in tumor cells, while exhibiting no toxicity in most normal cells. Since no potent apoptosis-inducing factor has been found yet in ameloblastoma, the present study was conducted. In the present study, expressions of TRAIL receptors, death receptor 4 (DR4) and DR5, were detected in all ameloblastoma tissues by immunohistochemistry as well as in AM-1 cells by immunofluorescence. By applying TRAIL in AM-1 cells, ameloblastoma cell line, for 24 h, we found that TRAIL cleaved caspase-8, -9 and –3, and lowered mitochondrial membrane potential (Δψm), and markedly induced apoptosis in AM-1 cells (46%). These results suggested that TRAIL is a potent apoptosis-inducing ligand in ameloblastoma.