RNA-sequencing identification and validation of genes differentially expressed in high-risk adenoma, advanced colorectal cancer, and normal controls.

Distinct gene expression patterns that occur during the adenoma-carcinoma sequence need to be determined to analyze the underlying mechanism in each step of colorectal cancer progression. Elucidation of biomarkers for colorectal polyps that harbor malignancy potential is important for prevention of colorectal cancer. Here, we use RNA sequencing to determine gene expression profile in patients with high-risk adenoma treated with endoscopic submucosal dissection by comparing with gene expression in patients with advanced colorectal cancer and normal controls. We collected 70 samples, which consisted of 27 colorectal polyps, 24 cancer tissues, and 19 normal colorectal mucosa. RNA sequencing was performed on an Illumina platform to select differentially expressed genes (DEGs) between colorectal polyps and cancer, polyps and controls, and cancer and normal controls. The Kyoto Gene and Genome Encyclopedia (KEGG) and gene ontology (GO) analysis, gene-concept network, GSEA, and a decision tree were used to evaluate the DEGs. We selected the most highly expressed genes in high-risk polyps and validated their expression using real-time PCR and immunohistochemistry. Compared to patients with colorectal cancer, 82 upregulated and 24 downregulated genes were detected in high-risk adenoma. In comparison with normal controls, 33 upregulated and 79 downregulated genes were found in high-risk adenoma. In total, six genes were retrieved as the highest and second highest expressed in advanced polyps and cancers among the three groups. Among the six genes, ANAX3 and CD44 expression in real-time PCR for validation was in good accordance with RNA sequencing. We identified differential expression of mRNAs among high-risk adenoma, advanced colorectal cancer, and normal controls, including that of CD44 and ANXA3, suggesting that this cluster of genes as a marker of high-risk colorectal adenoma.

The effect of adipose stem cell therapy on pulmonary fibrosis induced by repetitive intratracheal bleomycin in mice.

Adipose stem cells (ASCs) are detectable in the parenchyma and large airways of lungs after systemic administration, and ameliorate inflammatory infiltration and cell death in animal models of emphysema. We evaluated whether ASC treatment could attenuate lung fibrosis induced by repetitive intratracheal bleomycin administration. Male 8-week-old C57BL/6J mice (control group, bleomycin-only group, and bleomycin-plus-ASC group) were used. Eight biweekly doses of bleomycin were injected intratracheally via an intubation procedure at a dose of 0.04 units in a total volume of 100 µL of sterile saline. During the latter 2 months of the 4-month bleomycin exposure, human ASCs (3 × 10(5) cells) were administered repeatedly via intraperitoneal injection at the same time as bleomycin. Lung tissues were evaluated for histology, collagen content, TUNEL staining, and TGF-ß levels. Bronchoalveolar lavage (BAL) was performed for cell counting. Administrations of ASCs ameliorated the deleterious effects of repetitive intratracheal instillation of bleomycin, namely hyperplasia of Club cells (Clara cells) and cuboidal alveolar epithelial cells, infiltration of the perialveolar ducts by inflammatory cells, septal thickening, enlarged alveoli, and extensive fibrosis. Addition of ASC led to suppression of bleomycin-induced epithelial cell apoptosis and expression of TGF-ß. These results suggest a useful therapeutic effect of ASCs on pulmonary fibrosis induced by repetitive bleomycin administration. Further studies will be required to evaluate the efficacy of ASC therapy for the treatment of idiopathic pulmonary fibrosis.

Expression of moesin and CD44 is associated with poor prognosis in gastric adenocarcinoma.

AIMS: CD44 has been reported as a negative prognostic marker in gastric cancer. It interacts with moesin in epithelial-mesenchymal transition. To date, to our knowledge, there has been no clinical study dealing with the relationship between moesin and gastric adenocarcinoma. We analysed the expression of moesin and CD44 in gastric adenocarcinoma tissue, and correlations with clinicopathological factors. METHODS AND RESULTS: A retrospective analysis was made of 430 patients who had undergone gastrectomy at the Korea University Guro Hospital between 2002 and 2005 for gastric adenocarcinoma. Using tissue microarray and immunohistochemical staining, moesin expression was observed in 192 (44.7%) cases; it was associated significantly with poorly differentiated histology, invasion depth, lymph node metastasis, lymphatic invasion and advanced pathological TNM stage. CD44 expression was not correlated with clinicopathological features or moesin expression. Moesin expression was a strong predictor of lymph node metastasis in logistic regression analysis. Both moesin expression and CD44 expression were associated significantly with poor overall survival in univariate analysis. Furthermore, in multivariate analysis, moesin and CD44 were independent markers of poor prognosis, along with pathological TNM stage and older patient age. CONCLUSION: Moesin expression and CD44 expression might be useful markers of poor prognosis in gastric adenocarcinoma.

Successful salvage treatment of steroid-refractory bronchiolar COP with low-dose macrolides.

Cryptogenic organizing pneumonia (COP) is an idiopathic interstitial pneumonia characterized by fibroblastic tissues that occupy the lumina of alveoli and alveolar ducts or respiratory bronchioles. Although adequate doses and durations of glucocorticoids can improve its condition, COP is sometimes resistant to glucocorticoid therapy and is often lethal.Herein, a very rare case of 'bronchiolar COP' that was confined to the respiratory bronchioles is reported. This case indicates that macrolides may act as anti-inflammatory agents in patients with COP. Timely and precise pathological diagnosis may corroborate clinician diagnoses and eventually improve chances to overcome the disease.

A study of bioethical knowledge and perceptions in Korea.

This study assessed the knowledge and perception of human biological materials (HBM) and biorepositories among three study groups in South Korea. The relationship between the knowledge and the perception among different groups was also examined by using factor and regression analyses. In a self-reporting survey of 440 respondents, the expert group was found more likely to be knowledgeable and positively perceived than the others. Four factors emerged: Sale and Consent, Flexible Use, Self-Confidence, and Korean Bioethics and Biosafety Action restriction perception. The results indicate that those who are well aware of the existence of biobanks were more positively inclined to receive the Sale and Consent perception. As a result of the need for high quality HBMs and the use of appropriate sampling procedures for every aspect of the collection and use process, the biorepository community should pay attention to ethical, legal, and policy issues.

Anger, anger expression, and suicidal ideation in Korean adolescents.

This study described the levels of anger, anger expression, and suicidal ideation in Korean adolescents. Data from 18,752 adolescents were collected using a self-report questionnaire. Anger, anger expression, and suicidal ideation exhibited significant differences according to school level and gender. The group with higher anger and anger expression showed a higher average suicidal ideation score than that of the group with lower anger and anger expression, suggesting that school-based programs which alleviate anger may be needed to decrease suicidal ideation among Korean adolescents.

Expression of female sex hormone receptors and its relation to clinicopathological characteristics and prognosis of lung adenocarcinoma.

BACKGROUND: Adenocarcinoma (ADC) of the lung exhibits different clinicopathological characteristics in men and women. Recent studies have suggested that these differences originate from the expression of female sex hormone receptors in tumor cells. The aim of the present study was to evaluate the immunohistochemical expression of female sex hormone receptors in lung ADC and determine the expression patterns in patients with different clinicopathological characteristics. METHODS: A total of 84 patients with lung ADC who underwent surgical resection and/or core biopsy were recruited for the present study. Immunohistochemical staining was performed for estrogen receptor α (ERα), estrogen receptor ß (ERß), progesterone receptor (PR), epidermal growth factor receptor (EGFR), EGFR E746- A750 del, and EGFR L858R using tissue microarray. RESULTS: A total of 39 (46.4%) ERα-positive, 71 (84.5%) ERß-positive, and 46 (54.8%) PR-positive lung ADCs were identified. In addition, there were 81 (96.4%) EGFR-positive, 14 (16.7%) EGFR E746-A750 del-positive, and 34 (40.5%) EGFR L858R-positive cases. The expression of female sex hormone receptors was not significantly different in clinicopathologically different subsets of lung ADC. CONCLUSIONS: Expression of female sex hormone receptors is not associated with the prognosis and clinicopathological characteristics of patients with lung ADC.

Clinicopathological features and diagnostic methods of ALK fusion­positive non­small cell lung cancer in Korea.

Lung cancer is one of the most common malignancies and the leading cause of cancer­associated mortality in Korea. A significant amount of effort has been put into the development of new and more effective treatments and biological markers for the prediction of therapeutic responses, which has led to the identification of various genetic changes in cancer, that are the so­called 'growth drivers' of carcinogenesis. Certain genetic alterations have become new treatment targets, and it has been suggested that different mutations are associated with different clinicopathological characteristics and prognosis. The present study aimed to evaluate the status of the key 'driver' mutation anaplastic lymphoma kinase (ALK) fusion in Korean patients with non­small cell lung cancer (NSCLC) and its association with clinicopathological characteristics, including the presence of other genetic mutations. The present study also compared different methods for ALK fusion detection, including fluorescence in situ hybridization (FISH), immunohistochemistry (IHC) and next­generation sequencing (NGS) to evaluate which method is the most effective. A total of 482 patients with NSCLC who underwent ALK FISH analysis were evaluated for clinicopathological features, such as age, sex, smoking history, tumor stage, histological subtype, immunohistochemical profile, including ALK and EGFR mutation statuses, and survival. Some ALK FISH­positive and ­negative cancers were newly submitted to NGS analysis for DNA and RNA alterations. The ALK fusion­positive tumors were associated with a younger age, female patients, frequent nodal metastases, advanced stage and shorter survival. Comparing the results of ALK FISH, IHC and NGS analyses, it was concluded that in practice, ALK testing should better be diversified concerning FISH and IHC, and NGS analysis would be a good alternative to FISH, with an additional advantage of being able to concurrently detect different mutations.

High Paip1 Expression as a Potential Prognostic Marker in Hepatocellular Carcinoma.

BACKGROUND/AIM: Translation plays an important role in the carcinogenesis of various human tumors. Paip1 and eIF4A1 are translation-associated proteins that mediate the function of eukaryotic initiation factor 4F complex. This study aimed to analyse the relationship between the expression status of Paip1 and eIF4A1 and clinicopathologic features in hepatocellular carcinoma (HCC). MATERIALS AND METHODS: Immunohistochemical analysis was used to evaluate the expression status of Paip1 and eIF4A1. Two pathologists independently interpreted the immunostained slides. The prognostic value of Paip1 and eIF4A1 was evaluated by the Kaplan-Meier plotter. RESULTS: Among 173 HCC patients, 28 (16.1%) and 46 (26.6%) belonged in the Paip1 and eIF4A1 high-expression groups. High expression of Paip1 and eIF4A1 was associated with advanced TNM stage and more frequent vascular tumor invasion. Univariate analysis indicated that high Paip1 expression was associated with worse five-year overall survival (OS). Public dataset analysis by Kaplan-Meier plotter revealed that high mRNA expression of Paip1, and not of eIF4A1, was significantly associated with worse five-year OS and disease-free survival. CONCLUSION: Paip1 expression has a potential prognostic value in human HCC.

Expression and clinical significance of cell cycle regulatory proteins in gallbladder and extrahepatic bile duct cancer.

Disruption of cell cycle controls is a pathognomonic feature of all malignant cells. Therefore, we immunohistochemically investigated the relationship between cell cycle regulatory proteins and clinicopathologic features in order to identify the biomarkers related to the outcome of patients with biliary tract cancer (BTC). A cohort of paraffin-embedded specimens were selected from 36 patients, including 18 gallbladder and 18 extrahepatic bile duct cancers, who underwent curative or palliative surgical resection at Korea University Medical Center from June 1998 to December 2004. Tissue microarrays were used to investigate the immunohistochemical staining for p21, p27, p53, cyclin D1, bcl2, and Ki-67. Univariate and multivariate survival analyses were performed to determine the prognostic significance of each protein expression. Absence of p21 expression independently predicted poor outcome in all cases. Well-differentiated tumor was found to be an independent good prognostic factor in gallbladder cancer. Absence of p21 expression and moderately to poorly differentiated tumor were found to be an independent poor prognostic factor in patients with negative for neural invasion. Absence of p21 and bcl2 were found to be an independent poor prognostic factor in patients with no lymph node metastasis. Absence of p21 expression was a significant independent poor prognostic factor in BTC, partly in patients with biologically less aggressive phenotypes. This finding suggests that determination of p21 expression in surgically resected specimens may provide prognostic information in addition to conventional pathologic findings for patients with BTC, especially those who have biologically less aggressive phenotypes.

Bronchiolitis: classification, computed tomographic and histopathologic features, and radiologic approach.

The introduction of thin-section computed tomography (CT) has improved the diagnosis of bronchiolitis and contributed to establish bronchiolitis as an important part of pulmonary medicine. The histopathologic groups of bronchiolitis include cellular bronchiolitis, diffuse panbronchiolitis, follicular bronchiolitis, respiratory bronchiolitis, proliferative bronchiolitis, constrictive bronchiolitis, and mineral dust airway disease. Bronchiolitis can be readily identified and may lead to characteristic findings on thin-section CT. Various bronchiolitis manifests as one or a combination of 3 main patterns in thin-section CT: centrilobular nodules and branching densities, ground-glass attenuation and consolidation, and low attenuation and mosaic perfusion pattern.

High expression of DEK is associated with poor prognosis in hepatocellular carcinoma.

DEK is an oncogene that has been identified as part of the DEK-CAN fusion gene. DEK plays a role in carcinogenesis through WNT signaling and induces cell proliferation through cyclin-dependent kinase signaling. DEK overexpression has been reported in HCC, but the clinical significance is unclear. This study enrolled 221 cases of HCC. The expression of DEK protein was evaluated by immunohistochemical staining. Cdk4, cyclin D1, Wnt10b, E-cadherin, and ß-catenin were also immunohistochemically stained and analyzed for correlation. The association of clinicopathologic factors with DEK expression was analyzed. DEK expression was observed in 44.8% (99/221) of cases. DEK expression showed a statistical association with clinicopathologic factors, including Edmondson-Steiner grade, presence of vascular emboli, and multiplicity (p<0.05). Among the other IHC markers, the expression of cdk4 was correlated with DEK expression (p<0.05). Patients with high DEK expression showed a significantly lower overall survival rate (p=0.006). However, the disease-free survival rate did not differ significantly. In addition, in a Cox regression model analysis, DEK expression was an independent prognostic factor. In summary, high expression of DEK was observed in HCC and was associated with poor prognostic marker expression and poor prognosis.

Breast carcinomas expressing basal markers have poor clinical outcome regardless of estrogen receptor status.

To evaluate the clinical significance of gene expression-based classification and define the characteristic features of the new basal-like subtype, invasive breast carcinomas were divided into ER, HER2, basal-like and null subtypes by immunohistochemical analysis. A total of 401 invasive breast carcinomas were submitted to tissue microarray and stained with ER, HER2, EGFR, c-KIT and cytokeratin (CK) 5/6. The basal-like tumors, defined as positive for one or more basal markers but negative for both ER and HER2, comprised 18.5%. They were larger (p=0.041), showed higher grade (p<0.001), and more frequently expressed p53 (p=0.003). Expression of the basal marker itself showed negative prognostic effect, particularly in node-positive group. Even ER-positive patients had far shorter disease-free survival (DFS) when the tumor coexpressed one or more basal marker (p<0.001). Discrimination of basal-like subtype or tumors positive for basal markers may be clinically significant also in the treatment and prognosis of breast carcinomas.

Evolution of VX2 carcinoma in rabbit tibia: magnetic resonance imaging with pathologic correlation.

OBJECTIVE: To evaluate the evolution of a metastatic bone tumor model with MRI-pathology correlation. MATERIALS AND METHODS: VX2 carcinoma was implanted into the tibiae of 20 rabbits. The rabbits were divided into four groups of five (Groups I-IV). MRI was repeated at 1-week interval up to the fourth week, including sagittal T1-weighted image (T1WI), T2-weighted image (T2WI), gadolinium-enhanced fat-suppressed T1WI (GdT1WI), and diffusion-weighted image (DWI). Each group was sacrificed after the imaging, then histological examination for the tibiae with an implanted tumor was performed and MRI-pathologic correlation was done. RESULTS: On MRI-pathology correlation, the corresponding findings were as follows; low SI on T1WI, T2WI-tumor cells, fibrosis (1 week); central low SI on T1WI, T2WI, GdT1WI -tumor cells with fibrosis and necrosis; peripheral high SI on T2WI, DWI, GdT1WI-edema, fibrosis (2 weeks); heterogeneous SI with central low SI on T2WI, DWI-tumor cell nests with extensive necrosis, fibrosis; high SI on T2WI along periosteum-periosteal reaction; high SI around low SI and in bone marrow on T2WI, DWI, GdT1WI-edema, fibrosis; low SI on T1WI in surrounding bone marrow-tumor extension (3-4 weeks). CONCLUSION: The evolution of VX2 carcinoma model was well depicted on MR imaging. Necrosis and extent of tumor were best depicted on enhanced, fat-suppressed T1-weighted images. Heterogeneity of the tumor, peripheral edema, and fibrosis were represented well on T2-weighted images. Diffusion-weighted imaging could have a role in depicting necrosis in the evaluation of bone tumor.

Proteomic analysis of peripheral T-lymphocytes in patients with asthma.

BACKGROUND: Asthma is chronic airway inflammation that occurs together with reversible airway obstruction. T-lymphocytes play an important role in the pathogenesis of asthma. Proteomic technology has rapidly developed in the postgenomic era, and it is now widely accepted as a complementary technology to genetic profiling. We investigated the changes of proteins in T-lymphocytes of asthma patients by using standard proteome technology: two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS), and a database search. METHODS: The proteins of CD3+ T-lymphocytes were isolated from whole blood of six steroid-naive asthmatic patients and of six healthy volunteers. 2D-PAGE was performed and the silver-stained protein spots were comparatively analyzed between the asthma and control groups using an image analyzer. Some differentially expressed spots were identified by MALDI-TOF-MS and database search. The messenger RNA expressions of some identified proteins were examined by real-time polymerase chain reaction (RT-PCR). RESULTS: Thirteen protein spots in the T-lymphocytes of the asthmatic patients were increased and 12 spots were decreased compared to those of the normal subjects. Among the identified proteins, the increased expression of the messenger RNA of phosphodiesterase 4C and thioredoxin-2 and the decreased expression of the messenger RNA of glutathione S-transferase M3 were confirmed by RT-PCR in the asthmatic patients. CONCLUSIONS: Proteomic examination of the peripheral T-lymphocytes revealed some differentially expressed proteins in the asthmatic patients. The possibility of using the differentially expressed proteins as important biomarkers and therapeutic targets in asthma patients warrants further studies.

Correlated expression of erbB-3 with hormone receptor expression and favorable clinical outcome in invasive ductal carcinomas of the breast.

The prognostic impact of epidermal growth factor receptor (EGFR) family members in breast carcinoma was evaluated through the analysis of 378 cases of invasive ductal carcinoma to determine the relationship between EGFR family members and clinical outcome. It was found that 13.8% of the cases were positive for erbB-3. Expression of erbB-3 was inversely correlated with EGFR and erbB-4 expression (P = .012 and P = .046). Expression of erbB-3 was correlated with positive estrogen and progesterone receptor status and inversely correlated with histologic grade. Expression of erbB-3 was correlated with longer disease-free survival (DFS; P = .028). Within the erbB-3-expressing group, there was a tendency for the coexpressing group to have shorter DFS compared with the single-expressing group. This study revealed that erbB-3 expression was associated with better prognosis. In an era of personalized targeted therapy, erbB-3 expression and its coexpressive pattern with other EGFR family members could be an important determinant for therapeutic plans for breast cancer treatment.

Increased expression of matrix metalloproteinase 9 and tubulin-alpha in pulmonary sclerosing hemangioma.

Pulmonary sclerosing hemangioma (PSH) is relatively rare and is usually considered a benign tumor because of its slow growth and solitary characteristics. However, several cases with lymph node metastasis have been reported, and its pathogenesis has not been fully elucidated. Three sets of PSH specimens from the Korea Lung Tissue Bank, obtained with IRB approval, were analyzed through the construction of an oligo-microarray that contained about 32,000 genes. The resulting data were confirmed by real-time RT-PCR. Protein expression levels were checked by performing immunohistochemistry (IHC) and immunoblot analysis. In the 3 specimens of PSH tissues, 72 of the 32,000 genes were commonly found up-regulated and 290 were commonly found down-regulated as compared to non-tumor tissues from each patient. Paraffin-embedded tissues from 11 cases were used to confirm the expression of matrix metalloproteinase 9 (MMP-9) and tubulin-alpha proteins in the non-tumor and PSH tissues via IHC. In addition, the upregulation of protein expression was confirmed by immunoblot analysis. As expected, in all cases MMP-9 and tubulin-alpha were expressed at significantly higher levels in the PSH than in the non-tumor tissues. This is the first report on a study of the whole genome of PSH. Increased expression of MMP-9 could induce the metastatic ability of PSH and tubulin-alpha might be responsible for the sclerotic character of this disease. The results of this study will be useful in helping to understand and effectively manage patients suffering from PSH.

A Rare Case of Recurrent Metastatic Solid Pseudopapillary Neoplasm of the Pancreas.

A 61-year-old woman visited our hospital for bilateral multiple lung nodules and a mass in her thorax. She had a long history of multiple metastatic recurrences of solid pseudopapillary neoplasm (SPN); 24 years previously, the patient had undergone pylorus-preserving pancreaticoduodenectomy for a 9.9 × 8.6 cm mass in the pancreatic head. The tumor was diagnosed as an SPN. Nine years later, metastatic nodules were found on computed tomography in the patient's liver and peritoneum and were excised. She subsequently underwent an additional eight metastatectomy procedures in diverse organs. For the presented event, the lung nodules were removed. The prevalence of malignant SPN in the general population is 5%-15%. However, multiple metastatic recurrence of malignant SPN is rare; the lung is a particularly rare site of metastasis, found in only three cases in the literature. Here, we describe this exceptional case and provide a literature review.

Expression and prognostic significance of programmed death protein 1 and programmed death ligand-1, and cytotoxic T lymphocyte-associated molecule-4 in hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is one of the most common malignancies and causes of death worldwide. In this study, we assessed the correlation between clinicopathologic factors with programmed cell death protein 1 (PD-1) and programmed cell death ligand-1 (PD-L1), and cytotoxic T lymphocyte-associated molecule-4 (CTLA-4) expressions. Furthermore, we analyzed the prognostic significance of these proteins in a subgroup of patients. We retrospectively evaluated the PD-1, PD-L1, and CTLA-4 expressions in 294 HCC tissue microarray samples using immunohistochemistry. PD-1 and PD-L1 expressions were significant related to high CD8+ tumor-infiltrating lymphocytes (TILs) (r = 0.664, p < 0.001 and r = 0.149, p = 0.012). Only high Edmondson-Steiner grade was statistically related to high PD-1 expression. High PD-L1 expression was demonstrated as an independent poor prognostic factor for disease-free survival in addition to previous known factors, size >5 cm and serum albumin ≤3.5 g/dL in high CD8+ TILs group. We have demonstrated that the combined high expression of PD-L1 and CD8+ TIL is an important prognostic factor related to the immune checkpoint pathway in HCC and furthermore, there is a possibility that it could be used as a predictor of therapeutic response. Also, this result would be helpful in evaluating the applicable group of PD-1/PD-L1 blocking agent for HCC patients.