Noriyuki Nakano

Cyclophilin A expression and its prognostic significance in lung adenocarcinoma: CypA expression in lung adenocarcinoma

Cyclophilin A (CypA) has been reported to be upregulated in malignant tumors. CypA expression is thought to be associated with acquisition of tumor growth and anti‐apoptotic function. Although upregulation of CypA has been reported in lung adenocarcinoma, its clinicopathological significance and roles in malignant progression remain unclear. Here we investigated the implications of CypA expression for outcome in patients with lung adenocarcinoma. Lung adenocarcinoma specimens from 198 cases were selected and reclassified according to the World Health Organization classification (4th edition) and the Noguchi classification. CypA expression was assessed by immunohistochemistry, and the H‐score was calculated on the basis of intensity and proportion. The specificity of the antibody used was confirmed by Western blotting and the cut‐off point was determined from the ROC curve. Sixty‐seven cases (33.8%) had low CypA expression (CypA‐L group) and 131 (66.2%) had high CypA expression (CypA‐H group). Many cases of adenocarcinoma in situ were CypA‐L, and advanced adenocarcinomas tended to be classified as CypA‐H. Clinically, patients with CypA‐H tumors showed a significantly poorer prognosis than those with CypA‐L tumors. This is the first investigation of the implications of the CypA expression level in terms of the clinical characteristics of resected lung adenocarcinomas.

Developmental markers of ganglion cells in the enteric nervous system and their application for evaluation of Hirschsprung disease.

Hirschsprung disease (HSCR) is a congenital disease resulting from failure of neural crest‐derived ganglion cells to colonize the colon. Conventional diagnostic methods are insufficient for evaluating the ‘functional’ prognosis of HSCR. In order to elucidate the maturation of ganglion cells, 17 immunohistochemical markers were examined. We examined the digestive tracts of 2 human early delivery patients, 2 miniature swine fetuses, 4 little infants, 3 infants, 3 children, 6 adults, and 3 aged individuals. With increasing age, the labeling index (LI) for both calretinin and tyrosine hydroxylase (TH) increased, whereas that for SOX10 decreased. We then examined the ‘transitional zone’ of HSCR in 21 affected patients and 18 controls for these three markers. The LI of calretinin and TH were significantly lower than in the controls (median: 3.7 in HSCR and 8.2 in controls, P < 0.001, median: 27.9 in HSCR and 44.4 in controls, P < 0.001, respectively). In contrast, the LI for SOX10 showed no significant difference (median: 33.7 in HSCR and 29.2 in controls, P = 0.666) however, hierarchical cluster analysis was able to divide HSCR patients into two groups. These results suggest that immature ganglion cells are present in the transitional zone of HSCR, and that HSCR may have two different pathophysiological processes.

Novel non-alcoholic steatohepatitis model with histopathological and insulin-resistant features: A novel human-like NASH model

Although several non‐alcoholic steatohepatitis (NASH) models have been reported to date, few of these models fully reflect the histopathology and pathophysiology of human NASH. The aim of this study was to establish a novel NASH model by feeding a high‐fat (HF) diet and administering both carbon tetrachloride (CCl4) and the Liver X receptor agonist T0901317. Male C57BL/6J mice were divided into four groups (each n = 5): HF, HF + CCl4, HF + T0901317, and the novel NASH model (HF + CCl4 + T0901317). CCl4 (0.1 mL/kg) and T0901317 (2.5 mg/kg) were intraperitoneally administered four times and five times, respectively. The livers of the novel NASH model group presented a whitish colour. The serum levels of TNF‐α and IL‐6 were significantly increased in the novel NASH model group, and mice in this group exhibited histopathological features and insulin resistance reflective of NASH, i.e., macrovesicular hepatic steatosis, ballooning hepatocytes, Mallory‐Denk bodies, lobular inflammation and fibrosis. The novel NASH model group presented significantly upregulated expression levels of mRNAs related to lipogenesis, oxidative stress, fibrosis and steatosis and significantly downregulated expression levels of mRNAs related to triglyceride export. We successfully established a novel experimental NASH model that exhibits similar histopathology and pathophysiology to human NASH.

High expression of ovarian cancer immunoreactive antigen domain containing 2 (OCIAD2) is associated with poor prognosis in lung adenocarcinoma: OCIAD2 is prognostic marker of lung ad.

The clinicopathological implications of ovarian cancer immunoreactive antigen domain containing 2 (OCIAD2) in lung adenocarcinoma were investigated. The expression of OCIAD2 in 191 surgically resected lung adenocarcinomas was examined using immunohistochemistry. OCIAD2 expression was quantified using the H‐score and dichotomized as high or low. High OCIAD2 protein expression was significantly correlated with vascular invasion (P = 0.0018), lymphatic permeation (P = 0.049), T factor (P = 0.0024), and pathological stage (P = 0.0003). High OCIAD2 expression was significantly associated with poorer overall survival (OS) (n = 191, P = 0.0325). In peripheral‐type lung adenocarcinomas (n = 161), high OCIAD2 expression was significantly associated with both poorer OS (P = 0.0214) and poorer disease‐free survival (P = 0.0496). Adenocarcinoma in situ (AIS) and minimally invasive adenocarcinoma (MIA) showed weaker OCIAD2 expression than invasive adenocarcinoma. Among small adenocarcinomas measuring 2 cm or less in greatest dimension classified according to the Noguchis classification (n = 79), invasive adenocarcinomas showed significantly higher OCIAD2 expression than non‐invasive adenocarcinomas (P = 0.0007). Interestingly, OCIAD2 was expressed heterogeneously even within a tumor, and its expression was higher in areas of invasion than in areas of in situ spread. Our results suggest that OCIAD2 could be a useful prognostic biomarker of lung adenocarcinoma.

The prognostic significance of N-myc downregulated gene 1 in lung adenocarcinoma: NDRG1 in lung adenocarcinoma

It has been reported that N‐myc downstream regulated gene 1 (NDRG1) is related to the prognosis of non‐small cell lung cancer (NSCLC), and associated with c‐Myc degradation in NSCLC cell lines. However, the relationship of NDRG1 to prognosis or c‐Myc expression in lung adenocarcinoma has not been well clarified. The present study was designed to investigate the prognostic significance of NDRG1 and/or c‐Myc expression in lung adenocarcinoma using immunohistochemistry with a tissue microarray. We examined 184 lung adenocarcinomas and observed low expression of NDRG1 in adenocarcinoma in situ (AIS) and minimally invasive adenocarcinoma (MIA), whereas high expression of NDRG1 was seen in invasive adenocarcinoma. Each of the clinicopathological features except age was significantly correlated with NDRG1 expression. Kaplan‐Meier curves indicated that high expression of NDRG1 was significantly correlated with poor prognosis in comparison with low expression (log‐rank, P < 0.001). Univariate and multivariate analyses indicated that vascular invasion (P = 0.012), lymphatic permeation (P = 0.038), and NDRG1 expression (P = 0.026) were independent prognostic factors. Expression of NDRG1 and positivity for c‐Myc were significantly correlated (P = 0.005). These findings indicate that NDRG1 expression is associated with both prognosis and c‐Myc expression in lung adenocarcinoma.

Stratifin regulates stabilization of receptor tyrosine kinases via interaction with ubiquitin-specific protease 8 in lung adenocarcinoma

Previously we have reported that stratifin (SFN, 14-3-3 sigma) acts as a novel oncogene, accelerating the tumor initiation and progression of lung adenocarcinoma. Here, pull-down assay and LC-MS/MS analysis revealed that ubiquitin-specific protease 8 (USP8) specifically bound to SFN in lung adenocarcinoma cells. Both USP8 and SFN showed higher expression in human lung adenocarcinoma than in normal lung tissue, and USP8 expression was significantly correlated with SFN expression. Expression of SFN, but not of USP8, was associated with histological subtype, pathological stage, and poor prognosis. USP8 stabilizes receptor tyrosine kinases (RTKs) such as EGFR and MET by deubiquitination, contributing to the proliferative activity of many human cancers including non-small cell lung cancer. In vitro, USP8 binds to SFN and they co-localize at the early endosomes in lung adenocarcinoma cells. Moreover, USP8 or SFN knockdown leads to downregulation of tumor cellular proliferation and upregulation of apoptosis, p-EGFR or p-MET, which are related to the degradation pathway, and accumulation of ubiquitinated RTKs, leading to lysosomal degradation. Additionally, mutant USP8, which is unable to bind to SFN, reduces the expression of RTKs and p-STAT3. We also found that interaction with SFN is critical for USP8 to exert its autodeubiquitination function and avoid dephosphorylation by PP1. Our findings demonstrate that SFN enhances RTK stabilization through abnormal USP8 regulation in lung adenocarcinoma, suggesting that SFN could be a more suitable therapeutic target for lung adenocarcinoma than USP8.

Lung Carcinoma Representing Initially with Subacute Bilateral Isolated Hypoglossal Nerve Palsy: A case of atypical occipital condyle syndrome

We herein report the case of a 73‐year‐old man presenting bilateral hypoglossal nerve palsy as the initial symptom of metastatic lung cancer. The patient had subacute dysarthria and dysphagia caused by a disturbance in the protrusion of his tongue; He also had severe constant occipital pain, which prevented sleep. MRI showed a mass with contrast enhancement at the skull base. Squamous cell carcinoma was shown by bronchoscopic lung biopsy. In cases of hypoglossal nerve palsy with preceding severe ipsilateral occipital pain, metastatic cancer to the skull base should be considered as occipital condyle syndrome, even though the hypoglossal nerve palsy is bilateral or the patient has no history of malignant tumors. Occipital condyle syndrome with bilateral hypoglossal paralysis may be important as a sign of poor prognosis in cancer.

Heterotopic production of ceruloplasmin by lung adenocarcinoma is significantly correlated with prognosis

OBJECTIVES Ceruloplasmin (CP) is a well-known copper binding protein synthesized mainly in the liver, but its expression is known to be elevated in the serum of cancer patients and in malignant tumor cells. Lung cancer is the leading cause of cancer-related death worldwide, and adenocarcinoma is the main histological type of lung cancer. However, the role of CP in lung adenocarcinoma is still unclear. Here we examined and compared the expression of CP in various histological subtypes of lung adenocarcinoma and its correlation with clinicopathological parameters. MATERIALS AND METHODS We examined CP expression in lung adenocarcinoma samples and cell lines using quantitative real-time RT-PCR and Western blot analysis. Immunohistochemistry for CP was carried out using 196 specimens of lung adenocarcinoma. RESULTS CP expression was significantly higher in invasive adenocarcinoma than in adenocarcinoma in situ (AIS), and was significantly correlated with poorer outcome, pathological stage, pT, and pN. Multivariate analysis showed that CP expression was an independent prognostic factor for lung adenocarcinoma patients. Furthermore, Western blot analysis using protein extracted from lung adenocarcinoma cell lines revealed the secreted form of CP. CONCLUSION CP is produced heterotopically in lung adenocarcinoma cells and its expression is associated with tumor progression. In view of the presence of the secreted form of CP in tumor cells, CP may be a useful biomarker for lung adenocarcinoma.