Yu Usami

Intercellular adhesion molecule-1 (ICAM-1) expression correlates with oral cancer progression and induces macrophage/cancer cell adhesion

Intercellular adhesion molecule‐1 (ICAM‐1) is a transmembrane glycoprotein in the immunoglobulin superfamily, which plays an important role in cell adhesion and signal transduction. Although ICAM‐1 is believed to play a role in several malignancies, it is still uncertain whether or not ICAM‐1 expression contributes to cancer progression. In this study, we performed clinicopathological and cell biological analyses of ICAM‐1 expression in oral squamous cell carcinoma (SCC). First, we examined the ICAM‐1 expression in tongue SCC immunohistochemically, and revealed that ICAM‐1 was expressed predominantly at the invasive front area of tongue SCC. ICAM‐1 expression at the invasive front area was correlated with invasion, lymph node metastasis and increased blood and lymphatic vessel density of the tongue SCC. The relationship between ICAM‐1 expression and clinicopathological factors were consistent with the increased proliferation, invasion and cytokine‐production activities of ICAM‐1‐transfected SCC cells. Second, we analyzed the relationship between macrophages and ICAM‐1‐expressing tongue SCC cells because ICAM‐1 is known to act as a ligand for adhesion of immune cells. Increased ICAM‐1 expression in tongue SCC was correlated with increased macrophage infiltration within SCC nests. Moreover, macrophage/SCC‐cell adhesion through ICAM‐1 molecule was revealed using an in vitro cell adhesion and blockade assay. These findings indicate that ICAM‐1 plays an important role in tongue SCC progression, which may result from the SCC‐cell activity, angiogenic activity, lymphangiogenic activity and macrophage/SCC‐cell adhesion.

Sequential FDG-PET/CT after Neoadjuvant Chemotherapy is a Predictor of Histopathologic Response in Patients with Head and Neck Squamous Cell Carcinoma

PurposeTo evaluate whether 2-deoxy-2-[18F]fluoro-D-glucose positron emission tomography/computed tomography (FDG-PET/CT) more accurately predicts the histopathologic response to neoadjuvant chemotherapy (NAC) than magnetic resonance imaging (MRI) in patients with head and neck squamous cell carcinoma (HNSCC).ProceduresSixteen patients with HNSCC underwent FDG-PET/CT and MRI scans before and after one cycle of NAC, followed by surgical resection. The 26 surgically resected specimens of the 16 patients were analyzed. Decreases in maximum standardized uptake value (SUVmax) or in tumor maximum size (diametermax) were calculated, and their accuracies for the prediction of histopathologic response were evaluated.ResultsIn histopathologic responders (n = 7), percent decreases in SUVmax were significantly higher (P < 0.001) than in non-responders (n = 19). Applying a cut-off point of 55.5%, the histopathologic response could be predicted with a sensitivity and specificity of 86% and 95%, respectively.ConclusionFDG-PET/CT can predict histopathologic NAC responses with higher accuracy than MRI in HNSCC patients.

Differential expression of aquaporin 5 and aquaporin 3 in squamous cell carcinoma and adenoid cystic carcinoma

Aquaporins (AQPs) are a membrane protein family involved in the selective transport of water across cell membranes. Recent studies have reported the expression of AQP5 in several tumor types such as gastric, pulmonary, ovarian, pancreatic and colorectal cancer. We have previously reported the expression on tumor cells and the important role of AQP3 on cell growth in tongue cancer. However, little is known about the expression and precise role of AQP5 on squamous cell carcinoma (SCC) of the tongue. We investigated the expression of AQP5 and AQP3 in human oral SCC and adenoid cystic carcinoma (ACC). Overexpression of both AQP5 and AQP3 were immunohistochemically observed on tumor cells in SCC, whereas ACC cells were faintly stained with those antibodies against AQPs. Treatment with pan-AQP inhibitor or specific AQP5-siRNA showed inhibition of cell growth in SCC cell lines via the inhibition of integrins and the mitogen-activated protein kinase pathway. AQPs play important roles in cell growth in SCC rather than ACC.

Prognostic value of pretreatment 18F-fluorodeoxyglucose positron emission tomography/CT volume-based parameters in patients with oropharyngeal squamous cell carcinoma with known p16 and p53 status

The purpose of this study was to determine whether pretreatment 18F‐fluorodeoxyglucose‐positron emission tomography (18F‐FDG PET/CT) volume‐based parameters, such as metabolic tumor volume and total lesion glycolysis, add more prognostic information in patients with oropharyngeal squamous cell carcinoma (SCC).

Mild obesity reduces survival and adiponectin sensitivity in endotoxemic rats

BACKGROUND Recent meta-analyses have reported that critically ill patients with morbid obesity (body mass index >40 kg/m(2)) have poor outcomes, but the effects and mechanisms of action of mild obesity are still unclear. The purpose of this study was to evaluate the effect of mild obesity using a lard-based, high-fat diet (HFD) on pathologic conditions and the mechanisms of adiponectin action in endotoxemic rats. MATERIALS AND METHODS Male Wistar rats underwent HFD feeding for 4 wk and were killed at 0, 1.5, and 6 h after lipopolysaccharide (LPS) injection. Plasma levels of adiponectin, nitric oxide, and interleukin 6; messenger RNA expression of adiponectin receptors (AdipoR1 and AdipoR2) in the liver and the skeletal muscle; blood biochemical test results; and histology of the liver were analyzed. RESULTS HFD-fed rats had a lower survival rate (12.8% versus 85.2%) and lower plasma adiponectin levels after LPS injection (P < 0.01). Messenger RNA expression of adiponectin receptors in the liver, but not the skeletal muscle, also decreased in HFD-fed rats (P < 0.05). Tissue injury and oxidative stress in the liver and plasma inflammatory mediator levels increased, and worsened lipid metabolism abnormalities were noted. The findings indicated that HFD decreased the sensitivity of adiponectin and was associated with an increase in oxidative stress and inflammation, which finally resulted in worsened liver injury and poor survival rate after LPS injection. CONCLUSIONS Short-term, HFD-induced, mild obesity is harmful to the septic host, reduces adiponectin sensitivity, and could be the cause of worsening pathologic conditions.

Prognostic Impact of p16 and p53 Expression in Oropharyngeal Squamous Cell Carcinomas

BACKGROUNDS A p16 protein is known to be overexpressed in human papillomavirus-positive head and neck squamous cell carcinoma specimens. p53 is a tumor suppressor protein detectable by immunohistochemistry in carcinogen-associated head and neck squamous cell carcinoma as a result of gene mutations. The purpose of this study is to investigate the prognostic impact of p16 and p53 expression in oropharyngeal squamous cell carcinomas. METHODS We retrospectively examined the relationship between prognosis, and p16 and p53 expression levels of oropharyngeal squamous cell carcinoma specimens in 53 patients using immunohistochemistry. RESULTS Overall, 55% of patients were p16 positive and 45% p16 negative, while 28% were p53 positive and 72% p53 negative. The p16 status showed an inverse relationship with the p53 status. A survival analysis by p16 status, p53 status, Union for International Cancer Control stage and main treatment modality demonstrated that only p16 status was related to better prognosis in terms of overall survival and disease-specific survival (3-year overall survival, 87 vs. 62%, P = 0.02; 3-year disease-specific survival, 90 vs. 62%, P = 0.02). To evaluate the practical prognostic factors in oropharyngeal squamous cell carcinoma patients, we classified patients as either p16-positive or p53-negative oropharyngeal squamous cell carcinomas, representing human papillomavirus-related oropharyngeal squamous cell carcinoma with wild-type p53 or the remaining patients with p16-negative or p53-positive OPSCCs, respectively. The former group showed survival advantages in terms of overall survival and disease-specific survival by log-tank test compared with the latter group (3-year overall survival, 96 vs. 58%, P = 0.005; 3-year disease-specific survival, 96 vs. 63%, P = 0.02). CONCLUSIONS A group of patients who were p16 positive/p53 negative had better prognoses in terms of overall survival and disease-specific survival than that who were p16-positive alone.

Changes of hepatic lipid mediators associated with intake of high-fat diet for 12 weeks in endotoxemic rats using LC-ESI-MS/MS

BACKGROUND & AIMS It has recently been reported that anti-inflammatory lipid mediators are increased in the late phase of acute inflammation, whereas proinflammatory lipid mediators are regulated at the initiation of inflammation. The purpose of this study was to evaluate changes of hepatic lipid mediators due to high-fat diet (HFD) feeding in endotoxemic rats. METHODS Male Wistar rats were fed either HFD or control diet for 12 weeks, and were then killed 0, 1.5, and 6 h after lipopolysaccharide (LPS) injection. Analyses included lipidomics assessment of mediators using liquid chromatography-electrospray ionization/multi-stage mass spectrometry; measuring expression of hepatic polyunsaturated fatty acid (PUFA)-oxidizing enzyme, tumor necrosis factor (TNF)-α, interleukin (IL)-6, and inducible nitric oxide synthase mRNA levels; blood biochemical tests; and liver histology. RESULTS HFD feeding worsened liver injury, increased expression of TNF-α and IL-6 mRNA, and increased oxidative stress after LPS injection. PUFA-oxidizing enzymes were higher in HFD-fed rats after LPS injection. The proinflammatory prostaglandin (PG)E2 and thromboxane B2 were increased 1.5 h after LPS injection, and had decreased by 6 h in HFD-fed rats. In contrast, potent pro-resolving resolvins derived from eicosapentaenoic acid and docosahexaenoic acid were not detected, but anti-inflammatory epoxyeicosatrienoic acids, lipoxin A4, and 15-deoxy-PGJ2 were increased after LPS injection in HFD-fed rats. CONCLUSIONS HFD feeding for 12 weeks enhanced proinflammatory lipid mediators 1.5 h after LPS injection suggesting relation to liver injury.

A case of diffuse infiltrating gastrointestinal stromal tumor of sigmoid colon with perforation.

Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal neoplasms of the gastrointestinal tract, and typically present as discrete well‐circumscribed but non‐encapsulated tumor masses. In this report, we describe a case of colonic perforation caused by an unusual form of GIST. A 72‐year‐old Japanese woman presented to the emergency department with acute abdominal pain. Under the provisional diagnosis of sigmoid colon perforation, a laparoscopic sigmoidectomy was performed. Although the tumor mass was undetectable during the preoperative examination, a spindle cell lesion with a diffuse longitudinal growth pattern replacing the muscularis propria was revealed by microscopic examination. The spindle cell lesion was exposed at the perforation, suggesting a causal relationship between the lesion and the perforation. The spindle cell lesion was KIT‐positive and had a mutation in the C‐KIT gene at exon 11. We diagnosed it as diffuse infiltrating GIST. We consider that the lesion would be a cause of the colonic perforation, and emphasize the importance of accurate diagnosis of the lesion by histological, immunohistochemical and genetic examinations.

Neural hyperplasia in maxillary bone of multiple endocrine neoplasia type 2B patient

Multiple endocrine neoplasia (MEN) type 2B is the rarest and most aggressive form of MEN syndrome. MEN 2B patients manifest characteristic oral and facial features besides the neural crest cell-derived tumors, including medullary carcinoma, pheochromocytoma, mucosal neuroma, and ganglioneuromatosis of the gut. We report a case of MEN 2B diagnosed on the basis of the warning signs of mucosal neuroma and multiple neural hyperplasias in the maxillary bone resected during orthognathic surgery. A subsequent systemic examination under the pathologic diagnosis of neural lesions revealed medullary thyroid carcinoma, megacolon, thickened corneal nerves, and RET gene mutation, thus verifying the diagnosis of MEN 2B. An immunohistochemical study revealed an increased number of unmyelinated Schwann cells in the hyperplastic nerves. We suggest that intraosseous neural hyperplasia is a specific finding of the MEN 2B syndrome in addition to the known oral and facial manifestations.

Epigenetic upregulation of ARL4C, due to DNA hypomethylation in the 3'-untranslated region, promotes tumorigenesis of lung squamous cell carcinoma

ADP-ribosylation factor (ARF)-like 4c (ARL4C) expression, induced by a combination of Wnt/β-catenin and EGF/Ras signaling, has been demonstrated to form epithelial morphogenesis. ARL4C overexpression, due to Wnt/β-catenin and EGF/Ras signaling alterations, was involved in tumorigenesis. It was also reported that ARL4C expression correlates with DNA hypomethylation in the 3’-untranslated region (UTR) of ARL4C gene during lymphogenesis. The current study was conducted to investigate the expression and functions of ARL4C due to DNA hypomethylation in lung and tongue cancers. Immunohistochemical analyses of tissue specimens obtained from lung and tongue squamous cell carcinoma (SCC) patients revealed that ARL4C is not observed in non-tumor regions, but is strongly expressed at high frequencies in tumor lesions. Although inhibition of Wnt/β-catenin or Ras/MAP kinase signaling did not decrease ARL4C expression in NCI-H520 lung SCC cells, ARL4C DNA was clearly hypomethylated in the 3’-UTR. Ten-eleven translocation methylcytosine dioxygenase (TET) enzyme, which mediates DNA demethylation, was highly expressed in NCI-H520 cells. Knockout of TET family proteins (TET1-3) in NCI-H520 cells reduced 5-hydroxymethylcytosine (5hmC) levels and promoted DNA methylation in the 3’-UTR, leading to the decrease in ARL4C expression and ARL4C-mediated cellular migration. In tumor lesions of ARL4C-positive lung SCC, 5hmC was frequently detected and DNA methylation in the 3’-UTR of ARL4C gene was lower than in non-tumor regions, which were consistent with the Cancer Genome Atlas dataset. These results suggest that ARL4C is expressed due to hypomethylation in the 3’-UTR for certain types of cancers and that ARL4C methylation status is involved in cancer cell function.