Makoto Noguchi

Combined expression of p53, cyclin D1 and epidermal growth factor receptor improves estimation of prognosis in curatively resected oral cancer

p53, cyclin D1 and epidermal growth factor receptor (EGFR) are molecular markers that regulate the cell cycle or cell growth and play important roles in tumor development and progression. In this study, we examined the impact of immunohistochemical expression of these markers on tumor progression in 140 oral cancers. p53, cyclin D1 and EGFR were expressed in 64 cases (46%), 54 cases (39%) and 54 cases (39%), respectively, but there was no inter-relationship between any two of these markers. In the association of these markers with clinicopathological features, EGFR expression alone was significantly associated with poor differentiation (P=0.0008) and invasive growth pattern (P=0.0003). Any of these markers, including EGFR, had no significant impact on survival. Coexpression of all these markers, however, was significantly associated with invasive growth pattern (P=0.0149) and shortened survival (P=0.0181), and was a significant and independent unfavorable prognostic factor (P=0.0002), along with tumor size (P=0.0040), nodal metastasis (P=0.0137) and growth pattern (P=0.0017) in a multivariate analysis. Simultaneous coexpression of these markers in oral cancers might prove to be a useful indicator for identification of low- or high-risk patients.

Small interfering RNA-induced CHFR silencing sensitizes oral squamous cell cancer cells to microtubule inhibitors.

Alterations in the function of cell cycle checkpoints are frequently detected in oral squamous cell carcinomas (OSCCs), and are often associated with the sensitivity of the cancer cells to chemotherapeutic drugs. Recently, a mitotic checkpoint gene, Chfr, was shown to be inactivated by promoter methylation and point mutations in various human tumors. Here we show that the absence of its product, CHFR, is associated with mitotic checkpoint dysfunction, and that cancer cells lacking CHFR are sensitive to microtubule inhibitors. Checkpoint impairment appears to be caused by a prophase defect in this case, as OSCC cells lacking CHFR showed phosphorylation of histone H3 on Ser10 and translocation of cyclin B1 to the nucleus. When CHFR-deficient OSCC cells were treated with a microtubule inhibitor (docetaxel or paclitaxel), significant numbers of apoptotic cells were observed. Moreover, disruption of CHFR using small interfering RNA (siRNA) impaired the mitotic checkpoint, thereby reducing the ability of OSCC cells to arrest at G2/M phase and making them more sensitive to microtubule inhibitors. Our results suggest that CHFR could be a useful molecular target for chemotherapy.

Variations in the tensor veli palatini muscle with special reference to its origin and insertion.

Objective Previous research on the tensor veli palatini muscle (TVP) has produced conflicting descriptions of its functions and topographical relationships with other orofacial structures. The goal of this study was to describe the morphology of the TVP in a systematic and comprehensive manner. Methods One hundred nineteen sides of 77 human heads from donated cadavers were partially dissected under a binocular microscope. Histological examination of the hard tissue–muscle interfaces was also undertaken. Results There were two adjacent origins of the TVP: the cranial base origin (CB origin) and the auditory tube cartilage origin (AT origin). The CB origin always lay anterior to the AT origin and there was no septum or loose tissue between the two muscular laminae leading from these origins. The muscle fibers converged on a central tendinous plate in the muscle belly, which gradually became a common tendon that rounded the pterygoid hamulus before inserting into the palatine aponeurosis. Notably, secondary insertions were found on the maxillary tuber (33.6%) and/or in the submucosal tissue near the palatoglossal arch (37.8%). Maxillary insertions were almost exclusively associated with an AT origin that was wide as or wider than the CB origin. Histological observations confirmed that the hamulus acted purely as a pulley and suggested that a connecting band to the tensor tympani had no or few functions of an intermediate tendon. Conclusions The TVP appears to act as the dilator tubae and that this function can be maintained by preserving or reconstructing the maxillary insertion during push-back surgery, even if hamulotomy is necessary.

Constitutive Activation of Caspase-3 in Non-Apoptotic Oral Squamous Cell Carcinoma Cells

Background: Although caspase-3 is a key molecule for apoptosis induction, recent evidence has suggested its protumoral role in various human malignancies. The aim of the present study was to investigate the expression of cleaved caspase-3 (the active form of caspase-3) in both clinical samples and cell lines from oral squamous cell carcinomas (OSCCs) and elaborate on its contribution to the protumor role in oral cancer. Methods: The expression of cleaved caspase-3 was immunohistochemically evaluated in samples from 30 patients with OSCCs. The samples were either from biopsies or surgically-resected specimens with a mix of clinical stages and tumor site origins. The expression of cleaved caspase-3 was further examined in three OSCC cell lines. Results: In addition to apoptotic cancer cells, all the cases of OSCCs demonstrated a surprisingly positive expression of cleaved caspase-3. A diffuse, cytoplasmic pattern was particularly prominent in in situ carcinoma cells, invasive carcinoma cells, and metastatic cancer cells that lacked apoptotic morphology. On the other hand, non-neoplastic, normal epithelial cells were completely negative for cleaved caspase-3. In all the OSCC cell lines studied, cleaved caspase-3 was expressed in the cytoplasm and nucleus of cancer cells. Flow cytometric analysis also confirmed that the activation level of caspase-3 in non-apoptotic cancer cells was relatively lower than that in apoptotic cancer cells. Moreover, caspase-3 inhibition by caspase-3 specific inhibitor decreased the proliferation of OSCC cells. Conclusions: Because cleaved caspase-3 is selectively expressed in non-apoptotic OSCC cells and is associated with cell proliferation, these findings implicate caspase-3 signaling in promoting the progression of oral cancer.

Clinical significance of computed tomography assessment for third molar surgery.

Surgical extraction of the third molar is the most commonly performed surgical procedure in the clinical practice of oral surgery. Third molar surgery is warranted when there is inadequate space for eruption, malpositioning, or risk for cyst or odontogenic tumor formation. Preoperative assessment should include a detailed morphologic analysis of the third molar and its relationship to adjacent structures and surrounding tissues. Due to developments in medical engineering technology, computed tomography (CT) now plays a critical role in providing the clear images required for adequate assessment prior to third molar surgery. Removal of the maxillary third molar is associated with a risk for maxillary sinus perforation, whereas removal of the mandibular third molar can put patients at risk for a neurosensory deficit from damage to the lingual nerve or inferior alveolar nerve. Multiple factors, including demographic, anatomic, and treatment-related factors, influence the incidence of nerve injury during or following removal of the third molar. CT assessment of the third molar prior to surgery can identify some of these risk factors, such as the absence of cortication between the mandibular third molar and the inferior alveolar canal, prior to surgery to reduce the risk for nerve damage. This topic highlight presents an overview of the clinical significance of CT assessment in third molar surgery.

Afferent and efferent lymph-collecting vessels of the submandibular nodes with special reference to the lymphatic route passing through the mylohyoid muscle

Although metastasis of cancer in the oral region to the submandibular node is well described, there has been no anatomic representation of lymph vessels penetrating the oral floor and draining into the node.

Clinical significance of interstitial collagen deposition at the invading edge in oral cancer: Immunohistochemistry for type I collagen

BackgroundChanges in interstitial collagen in human oral cancer have not yet been fully studied. We examined the relationship between the degree of interstitial collagen deposition at the invading edge of the tumor, and the clinical and pathologic findings in oral squamous cell carcinoma. We also investigated the therapeutic implication of the changes in distribution patterns of collagen deposition by comparing biopsy specimens and surgical specimens.MethodsImmunohistochemical staining was performed by the streptavidin-biotin method using antibody against human type I collagen for visualizing interstitial collagen in 50 biopsy and 45 surgical specimens.ResultsCarcinomas with scanty interstitial collagen in biopsy specimens tended to have highly malignant characteristics. Large carcinomas with scanty deposition both in biopsy and surgical specimens were likely to have positive resection margins in spite of radical surgery.ConclusionImmunostaining patterns for type I collagen of oral squamous cell carcinomas can provide information of importance in determining safe resection margins.

Roles of intracellular and extracellular ROS formation in apoptosis induced by cold atmospheric helium plasma and X-irradiation in the presence of sulfasalazine

&NA; Sulfasalazine (SSZ) is a well‐known anti‐inflammatory drug and also an inhibitor of the cystine‐glutamate antiporter that is known to reduce intracellular glutathione (GSH) level and increase cellular oxidative stress, indicating its anti‐tumor potential. However, the combination of SSZ with other physical modalities remains unexplored. Here, the effects of SSZ on cold atmospheric helium plasma (He‐CAP), which produces approximately 24 x higher concentration of hydroxyl radicals (. OH) compared to X‐irradiation (IR) in aqueous solution, and on IR‐induced apoptosis in human leukemia Molt‐4 cells were studied to elucidate the mechanism of apoptosis enhancement. Both the Annexin V‐FITC/PI and DNA fragmentation assay revealed that pre‐treatment of cells with SSZ significantly enhanced He‐CAP and IR‐induced apoptosis. Similar enhancement was observed during the loss of mitochondrial membrane potential, intracellular Ca2+ ions, and mitochondria‐ and endoplasmic reticulum‐related proteins. The concentration of intracellular reactive oxygen species (ROS) was much higher in He‐CAP treated cells than in X‐irradiated cells. On the other hand, strong enhancement of Fas expression and caspase‐8 and ‐3 activities were only observed in X‐irradiated cells. It might be possible that the higher concentration of intracellular and extracellular ROS suppressed caspase activities and Fas expression in He‐CAP‐treated cells. Notably, pretreating the cells with an antioxidant N‐acetyl‐l‐cysteine (NAC) dramatically decreased apoptosis in cells treated by He‐CAP, but not by IR. These results suggest that IR‐induced apoptosis is due to specific and effective ROS distribution since intracellular ROS formation is marginal and the high production of ROS inside and outside of cells plays unique roles in He‐CAP induced apoptosis. We conclude that our data provides efficacy and mechanistic insights for SSZ, which might be helpful for establishing SSZ as a future sensitizer in He‐CAP or IR therapy for cancer. Graphical abstract Figure. No caption available. HighlightsGSH depleting agent SSZ enhances apoptosis in combination with He‐CAP and IR.SSZ, combined with He‐CAP, enhances apoptosis via the intrinsic apoptotic pathway.SSZ potentiates IR‐induced apoptosis via intrinsic and extrinsic apoptotic pathway.ER stress‐Ca2+ mediated pathway also contribute to the enhancement of apoptosis.Excess ROS generation by He‐CAP suppresses Fas, caspase‐8, and caspase‐3 activities.

Myeloid-derived suppressor cells in oral cancer

Results: Out of the total patients (1763), the number of women diagnosed with CIN I, CIN II, CIN III and carcinoma of cervix were 58 (3.3%), 2 (0.1%), 5 (0.3%) and 4 (0.2%), respectively. Severe cervicitis was detected in 1552 (88.8%) women while mild and moderate cervicitis were found in 47 (2.7%) and 164 (9.3%) patients. The results revealed that highest percentages of patients with CIN, moderate and severe cervicitis were diagnosed in women of 30-50 years old. While all patients with carcinoma of cervix were detected in women of ≥50 years old; more than 96% of CIN and all carcinoma cases were associated with severe cervicitinds.T across HLA and ABO barriers is now possible with pre-transplant desensitization for highly sensitized patients, but recipients of HLA incompatible allograft are at increased risk of rejection and graft loss. 745 renal transplant biopsies, including 380 protocol biopsies at 1, 3, 6 and 12 months post-transplant, from 129 recipients of HLA-incompatible kidney allograft, with follow up of more than 1 year (median 3 years) at the Johns Hopkins Hospital were analyzed. Biopsyproven rejection was associated with approximately 30% lower 5 year graft survival. Subclinical rejections were identified by protocol biopsies in a significant proportion of patients: 36% for cell mediated rejection (CMR) and 13% for antibody mediated rejection (AMR). Protocol biopsies detected glomerulitis, a feature of AMR and an important risk factor for development of chronic transplant glomerulopathy, in 41% of all biopsies performed in the first year post transplant. Transplant glomerulopathy within the first year post transplant is associated with worse graft survival and was observed in 22% of patients in this study. Transplant glomerulopathy at 1year was detected by protocol biopsies in approximately 75% of cases. Arteritis was noted in the graft of approximately 30% patients. Protocol biopsies detected about half of the arteritis in the renal grafts. Preliminary analysis of the data did not show significant association of arteritis with worse graft survival. Theobservation support the value of protocol biopsies performed serially after transplantation in revealing early subclinical lesions amenable to treatment, and improving graft survival.

Effects on preoperative therapy for squamous cell carcinoma of the lower alveolus and gingiva.

術前療法後に外科療法を施行した下顎歯肉扁平上皮癌一次症例57例を対象とした。対象症例の縮小率, 軟組織での組織学的術前療法効果および硬組織中での組織学的術前療法効果の関係について検討し, 以下の結果を得た。1.縮小率が高いほど予後が良好であった。2.縮小率が高い症例は, 大星・下里分類で著効例が多く認められた。3.術前療法による顎骨中の腫瘍の組織学的効果は, 軟組織中の所見とほぼ同様であった。4.X線所見がinvasive typeの症例では, 術前療法後, 骨中に生活癌細胞の残存が認められる傾向にあった。