Naruo Tokuyasu

ARPP protein is selectively expressed in renal oncocytoma, but rarely in renal cell carcinomas

We have recently isolated a gene, Ankyrin-repeated protein with a proline-rich region (ARPP), that is highly expressed in the skeletal and cardiac muscle. Our previous immunohistochemical analysis revealed that ARPP expression was augmented in rhabdomyosarcoma but scarcely detectable in leiomyosarcoma, showing that ARPP is a useful marker for rhabdomyosarcoma. In the present study, we generated the anti-ARPP monoclonal antibody, YAS11, immunoreactive with the N-terminal region (amino-acids residues 1–145) of the ARPP protein. Further, we immunohistochemically analyzed 100 renal tumors including 14 oncocytomas, and 86 renal cell carcinomas (RCCs). We found that ARPP was highly expressed in 12 of the 14 (85.7%) oncocytomas, but was detectable in only four of the 86 (4.7%) RCCs. Interestingly, ARPP was not detected in any of 11 chromophobe RCCs, suggesting that ARPP may be useful for differential diagnosis between oncocytoma and chromophobe RCC. Furthermore, we found that ARPP was selectively expressed in part of the distal renal tubule in normal kidney. Immunoelectron microscopy with anti-ARPP antibody revealed that ARPP was localized in mitochondria and nuclei in both the normal distal renal tubule and oncocytoma, suggesting that oncocytoma may be derived from the distal nephron, and probably from part of the distal renal tubule.

Minichromosome maintenance 2 (MCM2) immunoreactivity in stage III human gastric carcinoma: clinicopathological significance

BackgroundThe origin licensing factor minichromosome maintenance 2 (MCM2) has recently been identified as a critical regulator of proliferation in both normal and neoplastic cells. This study examined whether MCM2 expression was of prognostic relevance in patients with stage III gastric carcinoma and whether the expression of this marker showed any correlation with clinicopathological characteristics. In addition, we evaluated whether the expression of this proliferation marker was correlated with that of another marker, Ki-67, in gastric carcinoma.MethodsWe examined the immunohistochemical expression of MCM2, Ki-67, and p53 in 103 surgically removed stage III gastric carcinomas, which consisted of 60 intestinal-type and 43 diffuse-type carcinomas. The labeling indices (LIs) of MCM2 and Ki-67 in cancer cells were compared with clinicopathological characteristics, p53 expression, and overall survival rates.ResultsThe mean MCM2 and Ki-67 LIs were 69.1 ± 11.8% and 48.2 ± 14.5%, respectively, in the intestinal carcinomas, and 43.7 ± 9.9% and 24.9 ± 11.0%, respectively, in the diffuse carcinomas. The LIs of these proteins revealed no significant association with clinicopathological characteristics or with p53 expression in the carcinomas. Kaplan-Meier survival curves showed that, in the patients with diffuse carcinoma, those with higher MCM2 LIs had a poorer prognosis (P < 0.05), but the MCM2 LI was not correlated with prognosis for those with intestinal carcinoma (P = 0.25). Ki-67 expression had no significant correlation with prognosis in either intestinal-type or diffuse-type carcinomas. Multivariate Cox regression analysis confirmed that MCM2 was an independent prognostic factor in patients with diffuse carcinoma.ConclusionOur data suggest that MCM2 is a useful prognostic marker in patients stage III diffuse-type gastric carcinoma.

Expression of RUNX3 Protein in Human Esophageal Mucosa and Squamous Cell Carcinoma

Runt-related transcriptional factor gene 3 (RUNX3) belongs to the runt domain family of transcriptional factors that plays an important role during normal tissue development and in tumorigenesis in several organs. This study examined the expression of RUNX3 protein in human esophageal mucosa and squamous cell carcinoma in comparison with clinicopathological profiles. Western blot analysis and RT-PCR revealed that both RUNX3/P44 and P27, but not P46, were expressed in all three human esophageal squamous cell carcinoma (SCC) cell lines, as well as in three pairs of esophageal SCC cell lines and the corresponding nontumoral mucosa specimens. RUNX3 expression was shown in prickle and functional cell layer cells in normal esophageal mucosa. On the other hand, immunoreactivity was seen only in carcinoma cells around the cancer pearls. RUNX3 expression was significantly higher in the 19 well-differentiated SCCs than in the 56 moderately or 69 poorly differentiated SCCs (p < 0.01). The 3-year survival rate was significantly lower in the 29 patients with lower RUNX3 expression than in the 37 patients with higher expression (p = 0.0003). These results indicated that RUNX3 protein might play an important role in cellular differentiation in both esophageal mucosa and SCC. The expression correlated with the patients’ prolonged survival, implying a tumor suppressive effect in esophageal SCCs.

MAD1 (mitotic arrest deficiency 1) is a candidate for a tumor suppressor gene in human stomach

Mitotic arrest deficiency 1 (MAD1) is a component of the spindle checkpoint factors that monitor fidelity of chromosomal segregation. We previously confirmed that the level of MAD1 protein was decreased in gastric carcinoma compared with non-tumoral mucosa by conducting proteome-based analyses (Nishigaki R, Osaki M, Hiratsuka M, Toda T, Murakami K, Jeang KT, Ito H, Inoue T, Oshimura M, Proteomics 5:3205–3213, 29). In this study, an immunohistochemical analysis was performed to examine MAD1 expression histologically in gastric mucosa and tumor. MAD1 was detected in the supranuclear portion of normal epithelial, intestinal metaplasia, and adenoma cells, but its expression was not restricted to any specific area in carcinoma cells. Lower levels of expression were noted in 16 (47.1%) of 34 adenomas and in 52 (60.5%) of 86 carcinomas, whereas all normal mucosae and intestinal metaplasias were grouped into cases with higher level of expression. Moreover, the expression of MAD1 was significantly lower in advanced carcinomas than early carcinomas and in intestinal than diffuse type, respectively (P < 0.05). Exogenous expression of wild-type MAD1, but not the mutant MAD1, inhibited cell proliferation and resulted in G2/M accumulation in MKN-1, a gastric carcinoma cell line. Taken together, our findings suggest that the MAD1 gene could be a candidate tumor suppressor gene and that down-regulation of MAD1 expression contribute to tumorigenesis in human stomach.

Nicotine enhances the malignant potential of human pancreatic cancer cells via activation of atypical protein kinase C

BACKGROUND Pancreatic cancer (PC) is the most lethal malignancy among solid tumors, and the most common risk factor for its development is cigarette smoking. Atypical protein kinase C (aPKC) isozymes function in cell polarity, proliferation, and survival, and have also been implicated in carcinogenesis. However, the involvement of aPKC in PC progression and the effect of nicotine, a major component of cigarette smoke, on the biological activities of aPKC remain to be fully elucidated. METHODS We investigated the effects of nicotine on the proliferation, migration and invasion of the human PC cell lines Panc1 and BxPC3. We analyzed aPKC localization and activity by immunohistochemistry and in vitro kinase assays, respectively, to assess their involvement in the regulation of PC progression. Moreover, we examined the effect of nicotine on implanted peritoneal tumors of PC cells in mice. RESULTS Nicotine enhanced cell proliferation, migration and invasion in Panc1 and BxPC3 cells. In nicotine-treated PC cells, the aPKC was significantly activated. We also found that nicotine induced phosphatidylinositol 3-kinase (PI3K) signal activation, and a specific inhibitor of the nicotine acetylcholine receptor (nAChR) as well as knockdown of nAChR prevented nicotine-mediated Akt phosphorylation and aPKC activation. In a peritoneal dissemination model of PC, nicotine-treated mice had larger tumors and increased numbers of nodules. Immunohistochemistry showed enhanced expression levels of aPKC and phosphorylated Akt in nodules from nicotine-treated mice. CONCLUSIONS AND GENERAL SIGNIFICANCE Nicotine induces aberrant activation of aPKC via nAChR/PI3K signaling in PC cells, resulting in enhancement of cellular proliferation, migration and invasion.

Internal supravesical hernia repaired via the anterior approach alone: A case report

Highlights • Internal supravesical hernia is very rare.• Most previous reports performed closure of the hernial sac by open laparotomy.• We could successfully repair intraoperatively-diagnosed internal supravesical hernia by the anterior approach alone.

The attenuation value of preoperative computed tomography as a novel predictor for pancreatic fistula after pancreaticoduodenectomy

PurposePancreatic fistula (PF) is the most serious complication following pancreaticoduodenectomy (PD). This study was performed to identify new clinical factors that may predict the development of PF after PD to improve perioperative management.MethodsSeventy-five consecutive patients who underwent PD from 2012 to 2015 were evaluated. The patients’ perioperative data including the computed tomography (CT) parameters were collected. The minimum, maximum, and mean CT attenuation values (HUmin, HUmax, and HUmean, respectively) were extracted from the pancreatic parenchyma (≥ 100 pixels), and the standard deviation of these values (HUSD) was determined from the slice in which the superior mesenteric and splenic veins were merged. PF was defined as grade B or C according to the International Study Group for Pancreatic Fistula criteria.ResultsThe PF occurrence rate (grade B or C) was 25.3% in 75 patients. A multivariate analysis identified a larger HUSD (odds ratio 3.092; 95% CI 1.018–9.394) and higher amylase concentration in drainage fluid on postoperative day 1 (odds ratio 1.0001; 95% CI 1.00001–1.00022) as significant risk factors for PF.ConclusionsThe HUSD of preoperative CT attenuation values in the pancreatic parenchyma was found to be an independent predictor for PF after PD and it might therefore positively contribute to the perioperative management of PD.

Prognostic value of postoperative complication for early recurrence after curative resection of hepatocellular carcinoma

BACKGROUND Postoperative complications may adversely affect oncological outcomes. The aim of this study was to evaluate the impact of postoperative complications on early-phase recurrence after curative resection for hepatocellular carcinoma (HCC). METHODS We included 145 HCC patients who underwent initial and curative resection between January 2004 and December 2013. Postoperative complications of grade III or higher based on Clavien-Dindo classification were defined as clinically relevant postoperative complications. Recurrence within two years after hepatectomy was defined as early-phase recurrence. RESULTS Thirty-eight patients (26%) developed postoperative complications. The only predictive factor for postoperative complication was longer operative duration (P = 0.037). The disease-specific survival rate of patients with complication was lower than that of patients without complications (P = 0.015). Early-phase recurrence was observed in 20/38 (53%) patients who suffered postoperative complications and 36/107 (34%) patients with no complications, which was statistically significant (P = 0.039). Multivariate analysis identified four factors contributing to early-phase recurrence: high serum AFP level (P = 0.042), multiple tumors (P < 0.001), poor differentiation (P = 0.036) and presence of postoperative complication (P = 0.039). CONCLUSIONS Postoperative complication is an independent prognostic factor for early-phase recurrence after curative resection of HCC. Close observation of patients with postoperative complications may be a necessary treatment strategy for HCC.

The Combination of Neutrophil–to–lymphocyte Ratio and Serum Carbohydrate Antigen 19-9 Level as a Prognostic Indicator in Patients with Recurrent Pancreatic Cancer

Background/Aim: We retrospectively investigated the relationship between prognosis and combined neutrophil–to–lymphocyte ratio (NLR) and serum carbohydrate antigen 19-9 (CA19-9) levels in patients with recurrent pancreatic cancer. Patients and Methods: We enrolled 66 patients whose pancreatic cancer recurred. Results: Based on ROC analysis results, the patients were divided into NLRHigh (NLR ≥1.69) or NLRLow (NLR <1.69), and into CA19-9High (CA19-9 ≥107.95 U/ml) or CA19-9Low (CA19-9 <107.95 U/ml). When the patients were grouped by combined NLR and CA19-9, their 2-year survival rates were NLRLow/CA19-9Low: 58.7%; NLRLow/CA19-9High or NLRHigh/CA19-9Low (grouped together): 11.2%; and NLRHigh/CA19-9High: 0% (p<0.0001). Finally, in multivariate analysis, the combination of NLR and serum CA19-9 level was an independent prognostic factor in patients with recurrent pancreatic cancer. Conclusion: The combination of NLR and serum CA19-9 level is a useful prognostic indicator for recurrent pancreatic cancer.

Pilot Study of Probe-based Confocal Laser Endomicroscopy with Fluorescein-dripping Method for Liver Tumors

Background/Aim: Probe-based confocal laser endomicroscopy (pCLE) is a novel diagnostic technique that can provide real-time imaging of tissues at the cellular level. In this study, pCLE was applied to liver tumors and its diagnostic value was evaluated. Patients and Methods: Ten patients who underwent hepatectomy for liver tumors were evaluated with pCLE from February to May 2017. Immediately after liver resection, pCLE evaluation was performed in the operating room as an ex vivo study. The newly-adopted “fluorescein-dripping method” was used for the observation. Results: First, the optimal fluorescein exposure time for the surface of liver was defined in the fluorescein-dripping method. Next, the distinctive findings in the cancerous region were investigated. The characteristic appearance of irregular arrangements of concentrated cells under fluorescein dripping was observed in six of seven hepatocellular carcinoma (HCC) tumors. Conclusion: In all HCC specimens, discrimination of the cancerous region from normal liver was possible with pCLE.