Hypermethylation of TMEM240 predicts poor hormone therapy response and disease progression in breast cancer.

BACKGROUND: Approximately 25% of patients with early-stage breast cancer experience cancer progression throughout the disease course. Alterations in TMEM240 in breast cancer were identified and investigated to monitor treatment response and disease progression. METHODS: Circulating methylated TMEM240 in the plasma of breast cancer patients was used to monitor treatment response and disease progression. The Cancer Genome Atlas (TCGA) data in Western countries and Illumina methylation arrays in Taiwanese breast cancer patients were used to identify novel hypermethylated CpG sites and genes related to poor hormone therapy response. Quantitative methylation-specific PCR (QMSP), real-time reverse transcription PCR, and immunohistochemical analyses were performed to measure DNA methylation and mRNA and protein expression levels in 394 samples from Taiwanese and Korean breast cancer patients. TMEM240 gene manipulation, viability, migration assays, RNA-seq, and MetaCore were performed to determine its biological functions and relationship to hormone drug treatment response in breast cancer cells. RESULTS: Aberrant methylated TMEM240 was identified in breast cancer patients with poor hormone therapy response using genome-wide methylation analysis in the Taiwan and TCGA breast cancer cohorts. A cell model showed that TMEM240, which is localized to the cell membrane and cytoplasm, represses breast cancer cell proliferation and migration and regulates the expression levels of enzymes involved in estrone and estradiol metabolism. TMEM240 protein expression was observed in normal breast tissues but was not detected in 88.2% (67/76) of breast tumors and in 90.0% (9/10) of metastatic tumors from breast cancer patients. QMSP revealed that in 54.5% (55/101) of Taiwanese breast cancer patients, the methylation level of TMEM240 was at least twofold higher in tumor tissues than in matched normal breast tissues. Patients with hypermethylation of TMEM240 had poor 10-year overall survival (p = 0.003) and poor treatment response, especially hormone therapy response (p < 0.001). Circulating methylated TMEM240 dramatically and gradually decreased and then diminished in patients without disease progression, whereas it returned and its levels in plasma rose again in patients with disease progression. Prediction of disease progression based on circulating methylated TMEM240 was found to have 87.5% sensitivity, 93.1% specificity, and 90.2% accuracy. CONCLUSIONS: Hypermethylation of TMEM240 is a potential biomarker for treatment response and disease progression monitoring in breast cancer.

BHLHE22 Expression Is Associated with a Proinflammatory Immune Microenvironment and Confers a Favorable Prognosis in Endometrial Cancer.

Endometrial cancer (EC) rates are rising annually. Additional prediction markers need to be evaluated because only 10-20% of EC cases show an objective response to immune-checkpoint inhibitors (ICIs). Our previous methylomic study found that BHLHE22 is hypermethylated in EC tissues and can be detected using a Pap-smear sample. BHLHE22, a basic helix loop helix transcription factor family member, is known as a transcriptional repressor and is involved in cell differentiation. However, the role of BHLHE22 in EC remains poorly understood. Herein, we analyzed BHLHE22 expression in 54 paired cancer and normal endometrial tissue samples, and confirmed with databases (TCGA, GTEx, and human protein atlas). We found that BHLHE22 protein expression was significantly downregulated in EC compared with normal endometrium. High BHLHE22 expression was associated with microsatellite-instable subtype, endometrioid type, grade, and age. It showed a significant favorable survival. BHLHE22 overexpression inhibited the proliferation and migration of EC cells. Functional enrichment analysis showed that BHLHE22 was significantly associated with immune-related pathways. Furthermore, BHLHE22 was positively correlated with proinflammatory leukocyte infiltration and expression of chemokine genes in EC. In conclusion, BHLHE22 regulates immune-related pathways and modulates the immune microenvironment of EC.

Epigenomic Analysis Reveals the KCNK9 Potassium Channel as a Potential Therapeutic Target for Adenomyosis.

Adenomyosis is linked to dysmenorrhea and infertility. The pathogenesis of adenomyosis remains unclear, and little is known of the genetic and epigenetic changes in the eutopic endometrium in adenomyosis, which may predispose patients to the invasion and migration of endometrial tissues into the myometrium. Transcriptome studies have identified genes related to various cell behaviors but no targets for therapeutic intervention. The epigenetics of the eutopic endometrium in adenomyosis have rarely been investigated. Endometrial tissue was obtained from premenopausal women with (n = 32) or without adenomyosis (n = 17) who underwent hysterectomy aged 34-57 years at a tertiary hospital. The methylome and transcriptome were assessed by using a Methylation 450 K BeadChip array and Affymetrix expression microarray. Protein expression was examined by immunohistochemistry. Differential methylation analysis revealed 53 lowly methylated genes and 176 highly methylated genes with consistent gene expression in adenomyosis, including three genes encoding potassium ion channels. High expression of KCNK9 in the eutopic and ectopic endometria in patients with adenomyosis but not in normal controls was observed. Hormone-free, antibody-based KCNK9 targeting is a potential therapeutic strategy for adenomyosis-related dysmenorrhea, menorrhagia, and infertility.

Distinct methylation profile of mucinous ovarian carcinoma reveals susceptibility to proteasome inhibitors.

Mucinous type of epithelial ovarian cancer (MuOC) is a unique subtype with a poor survival outcome in recurrent and advanced stages. The role of type-specific epigenomics and its clinical significance remains uncertain. We analyzed the methylomic profiles of 6 benign mucinous adenomas, 24 MuOCs, 103 serous type of epithelial ovarian cancers (SeOCs) and 337 nonepithelial ovarian cancers. MuOC and SeOC exhibited distinct DNA methylation profiles comprising 101 genes, 81 of which exhibited low methylation in MuOC and were associated with the response to glucocorticoid, ATP hydrolysis-coupled proton transport, proteolysis involved in the cellular protein catabolic process and ion transmembrane transport. Hierarchical clustering analysis showed that the profiles of MuOC were similar to colorectal adenocarcinoma and stomach adenocarcinoma. Genetic interaction network analysis of differentially methylated genes in MuOC showed a dominant network module is the proteasome subunit beta (PSMB) family. Combined functional module and methylation analysis identified PSMB8 as a candidate marker for MuOC. Immunohistochemical staining of PSMB8 used to validate in 94 samples of ovarian tumors (mucinous adenoma, MuOC or SeOC) and 62 samples of gastrointestinal cancer. PSMB8 was commonly expressed in MuOC and gastrointestinal cancer samples, predominantly as strong cytoplasmic and occasionally weak nuclei staining, but was not expressed in SeOC samples. Carfilzomib, a second-generation proteasome inhibitor, suppressed MuOC cell growth in vitro. This study unveiled a mucinous-type-specific methylation profile and suggests the potential use of a proteasome inhibitor to treat MuOC.

GATA3 as a master regulator and therapeutic target in ovarian high-grade serous carcinoma stem cells.

Ovarian high-grade serous carcinoma (HGSC) is the most lethal gynecological malignancy. Prevailing evidences suggest that drug resistance and recurrence of ovarian HGSC are caused by the presence of cancer stem cells. Therefore, targeting cancer stems is appealing, however, all attempts to date, have failed. To circumvent this limit, we analyzed differential transcriptomes at early differentiation of ovarian HGSC stem cells and identified the developmental transcription factor GATA3 as highly expressed in stem, compared to progenitor cells. GATA3 expression associates with poor prognosis of ovarian HGSC patients, and was found to recruit the histone H3, lysine 27 (H3K27) demethylase, UTX, activate stemness markers, and promote stem-like phenotypes in ovarian HGSC cell lines. Targeting UTX by its inhibitor, GSKJ4, impeded GATA3-driven stemness phenotypes, and enhanced apoptosis of GATA3-expressing cancer cells. Combinations of gemcitabine or paclitaxel with GSKJ4, resulted in a synergistic cytotoxic effect. Our findings provide evidence for a new role for GATA3 in ovarian HGSC stemness, and demonstrate that GATA3 may serve as a biomarker for precision epigenetic therapy in the future.

Epigenetic loss of heparan sulfate 3-O-sulfation sensitizes ovarian carcinoma to oncogenic signals and predicts prognosis.

Precision medicine requires markers for therapeutic guidance. The purpose of this study was to determine whether epithelial ovarian cancer (EOC) epigenetics can lead to the identification of biomarkers for precision medicine. Through integrative methylomics, we discovered and validated the epigenetic signature of NEFH and HS3ST2 as an independent prognostic factor for type II EOC in our dataset (n = 84), and two independent methylomics datasets (total n = 467). Integrated transcriptomics dataset (n = 1147) and tissue microarrays (n = 54) of HS3ST2 also related to high-methylation statuses and the EOC prognosis. Mechanistic explorations of HS3ST2 have assessed responses to oncogenic stimulations such as IL-6, EGF, and FGF2 in cancer cells. The combination of HS3ST2 and various oncogenic ligands also confers the worse outcome. 3-O-sulfation of heparan sulfate by HS3ST2 makes ovarian cancer cells intrinsically sensitive to oncogenic signals, which sheds new light on the application of HS3ST2 as a companion diagnostic for targeted therapy using kinase inhibitors or therapeutic antibodies.

Active DNA Demethylase, TET1, Increases Oxidative Phosphorylation and Sensitizes Ovarian Cancer Stem Cells to Mitochondrial Complex I Inhibitor.

Ten-eleven translocation 1 (TET1) is a methylcytosine dioxygenase involved in active DNA demethylation. In our previous study, we demonstrated that TET1 reprogrammed the ovarian cancer epigenome, increased stem properties, and activated various regulatory networks, including metabolic networks. However, the role of TET1 in cancer metabolism remains poorly understood. Herein, we uncovered a demethylated metabolic gene network, especially oxidative phosphorylation (OXPHOS). Contrary to the concept of the Warburg effect in cancer cells, TET1 increased energy production mainly using OXPHOS rather than using glycolysis. Notably, TET1 increased the mitochondrial mass and DNA copy number. TET1 also activated mitochondrial biogenesis genes and adenosine triphosphate production. However, the reactive oxygen species levels were surprisingly decreased. In addition, TET1 increased the basal and maximal respiratory capacities. In an analysis of tricarboxylic acid cycle metabolites, TET1 increased the levels of α-ketoglutarate, which is a coenzyme of TET1 dioxygenase and may provide a positive feedback loop to modify the epigenomic landscape. TET1 also increased the mitochondrial complex I activity. Moreover, the mitochondrial complex I inhibitor, which had synergistic effects with the casein kinase 2 inhibitor, affected ovarian cancer growth. Altogether, TET1-reprogrammed ovarian cancer stem cells shifted the energy source to OXPHOS, which suggested that metabolic intervention might be a novel strategy for ovarian cancer treatment.

Gastrointestinal quality of life following bariatric surgery in Asian patients.

BACKGROUND/AIMS: Bariatric surgery is the only proven method that produces sustained weight loss. We aimed to investigate the Gastrointestinal Quality of Life Index (GIQLI) differences between obese patients following laparoscopic mini-gastric bypass (LMGB), laparoscopic adjustable gastric banding (LAGB) and laparoscopic sleeve gastrectomy (LSG) in this study. METHODOLOGY: From December 2005 to December 2007, we enrolled 152 patients who received bariatric surgery, including 41 men and 111 women, mean age 32.6±9.4 years and mean BMI 37.4±7.9kg/m2 (range 32.0-64.9). Clinical characteristics and quality of life were analyzed. RESULTS: One year after bariatric surgery, the mean general score of GIQLI improved significantly (p=0.000). All patients had improvement in three domains of the questionnaire (social function, physical status and emotional status) but not in gastrointestinal symptoms. The preoperative general score was 105.9±15.4 points in LMGB group, 110.9±14.8 points in LAGB group and 99.0±19.8 points in LSG group, respectively. Despite a significant difference between three groups regarding preoperative GIQLI scores (p=0.001), the 1-year results failed to show any significant difference in a comparison of postoperative GIQLI scores (p=0.082). CONCLUSIONS: In conclusion, our study has demonstrated significant improvement in quality of life 1-year after laparoscopic bariatric surgery. The improvement of GIQLI scores in three domains of social function, physical status and emotional status can be offered to obese patients before surgery.

Survivin-mediated therapeutic efficacy of gemcitabine through glucose-regulated protein 78 in hepatocellular carcinoma.

BACKGROUND: Survivin is an antiapoptotic molecule that is widely expressed in cancers, including hepatocellular carcinoma (HCC). Survivin has become a general therapeutic target for cancers because of its selective overexpression in a majority of tumors. However, little is known regarding the effect of survivin expression in combination with gemcitabine on HCC. METHODS: We generated survivin knockdown cells (survivin-KD) via a short interfering RNA (siRNA) technique. The antiproliferation effects of gemcitabine were determined by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, TUNEL (terminal deoxynucleotidyl transferase dUTP nick-end labeling) assay, and cell cycle evaluation. RESULTS: According to the MTT assay, we found that survivin-KD cells were more sensitive than parental cells and scrambled control cells to gemcitabine treatment. The apoptotic cell population increased in survivin-KD cells that were treated with gemcitabine in comparison to scrambled control cells, as observed by the cell cycle distribution and TUNEL assays. We found that survivin knockdown resulted in a reduction of glucose-regulated protein 78 (GRP78), which may be responsible for the observed increased survivin-KD cell sensitivity to gemcitabine. CONCLUSIONS: We conclude that survivin knockdown may contribute to a therapeutic effect of gemcitabine through GRP78 on HCC cells.

Roles of hepatic progenitor cells activation, ductular reaction proliferation and Notch signaling in morbid obesity.

BACKGROUND/AIMS: Hepatic progenitor cells (HPCs) activation, proliferative ductular reaction (DR), replicative arrest and Notch signaling have been demonstrated in a variety of human liver diseases. The relationships are poorly understood in morbid obesity. We investigated factors responsible for the HPCs/DR, replicative arrest and Notch signaling in non-NASH and NASH groups. METHODOLOGY: Cytokeratin 7 (and 19), p21, CD34, Ki67 and different Notch receptors and ligands immunohistochemical stained biopsies from morbid obese patients with non-NASH (n=10) and NASH (n=25) were studied. These results were correlated with clinicopathological variables. RESULTS: NASH patients presented with abnormal liver function tests and had higher HbA1c percentage. Strong association between HPCs and DR was seen (r=0.785, p<0.000). BMI, interface activity and replicative arrest were associated with HPCs expansion and DR in NASH patients. A strong association between CD34 with HPCs and DR was found in non-NASH patients. In NASH group, Notch 3 was important in bile ductular proliferation; whereas Notch 4 was associated with sinusoidal neovessels proliferation and Kupffer cell activation. CONCLUSIONS: HPCs and DR played an important role in hepatic regeneration in fatty liver disease of morbid obesity. An altered replication pathway in NASH promotes HPCs activation and DR. Notch-3 and Notch-4 were significantly different between non-NASH and NASH groups.

Predictors of anemia after bariatric surgery using multivariate adaptive regression splines.

BACKGROUND/AIMS: Anemia is the most common nutritional deficiency after bariatric surgery. The predictors of anemia have not been clearly identified. This issue is useful for selecting an appropriate surgery procedure for morbid obesity. METHODOLOGY: From December 2000 to October 2007, a retrospective study of 442 obese patients after bariatric surgery with two years' follow-up data was conducted. Anemia was defined by hemoglobin (Hb) under 13mg/dL in male and 11.5mg/dL in female. We analyzed the clinical information and laboratory data during the initial evaluation of patients referred to bariatric surgery for predictors of anemia development after surgery. All data were analyzed by using multivariate adaptive regression splines (MARS) method. RESULTS: Of the patients, the mean age was 30.8±8.6 years; mean BMI was 40.7±7.8kg/m2 and preoperative mean hemoglobin (Hb) was 13.7±1.5g/ dL. The prevalence of anemia increased from preoperatively 5.4% to 38.0% two years after surgery. Mean Hb was significantly lower in patients receiving gastric bypass than in restrictive type surgery (11.9mg/dL vs. 13.1mg/dL, p=0.040) two years after surgery. Besides, the preoperative optimal value of hemoglobin to predict future anemia in MARS model is 15.6mg/dL. CONCLUSIONS: The prevalence of anemia increased to 38.0% two years after bariatric surgery. We obtained an optimal preoperative value of hemoglobin 15.6mg/dL to predict postoperative anemia, which was important in preoperative assessment for bariatric surgery. Patients undergone gastric bypass surgery developed more severe anemia than gastric banding or sleeve gastrectomy.

Hepatosplenic actinomycosis in an immunocompetent patient.

Hepatosplenic abscess caused by Actinomyces is rare and often misdiagnosed as malignancy. Herein, we report a case of hepatosplenic actinomycosis in a 37-year-old immunocompetent man with a 2-month clinical history of intermittent fever and upper left abdominal pain. Physical examination revealed a mildly ill-appearing man with a low-grade fever (38°C) and upper left quadrant abdominal tenderness. Abdominal sonographic examination showed the presence of a 6.3 cm × 6.5 cm heterogeneous abscess with a hypoechoic center and honeycomb appearance in an enlarged spleen (8 cm × 5 cm). Computerized tomography of the abdomen revealed a multiloculated splenic lesion, and laparotomy showed multiple hepatic nodules and a splenic abscess. Histopathological examination of the biopsy revealed filamentous branching bacilli and sulfur granules in the hepatosplenic abscess. The patient successfully underwent splenectomy accompanied by intravenous and oral penicillin treatment. Proper and prompt diagnosis of hepatosplenic actinomycosis is important because the therapeutic plan and prognosis of this pathogen are quite different from other microorganisms and malignancies.

Association of immunohistochemical profiles with histotypes in endometrial carcinomas.

OBJECTIVE: Although a large number of endometrial cancer patients are cured with surgery alone, there are significant numbers of patients with more aggressive variants of endometrial carcinoma for whom the prognosis remains poor. We investigated the effects of prevalence, histotypes, and immunohistochemical profiles on prognostic value in a hospital-based population. MATERIALS AND METHODS: A retrospective study of surgically resected primary endometrial carcinoma was included. Immunohistochemical stains were performed on formalin-fixed paraffin-embedded tissue microarray sections for ß-Catenin, estrogen receptor (ER), progesterone receptor (PR), HER-2, MLH1, MSH2, MSH6, PMS2, and p53. RESULTS: Loss of mismatch repair expression was detected in 25.4% of samples (29/114, mean age 57 years) of the tumors. The following loss of expression was observed in patients: MLH1/PMS2 in 16.6% of patients, MSH6 in 7.0% of patients, MLH1 in 0.9% of patients, and MSH6/PMS2/MLH1 in 0.9% of patients. Immunohistochemistry of p53 was analyzed for 111 patients. A total of 13 patients (11.7%, mean age 64 years) had p53-abnormal expression (absent, cytoplasmic or diffuse strong positive patterns), and more than half (9/13, 69.2%) had endometrioid histotype. Abnormalities in p53 were significantly associated with histotype (p = 0.001), advanced tumor stage (p = 0.038), death of disease (p = 0.002), PR percentage (p = 0.002), and HER-2 expression (p = 0.018). Immunohistochemical nuclear localization of ß-Catenin was detected in 7.1% of the cohort. The combination of p53 and nuclear ß-Catenin expressions was not significantly predictive of disease-free or overall survival. CONCLUSION: The results of this study are useful for management of endometrial cancer in patients with DNA mismatch repair, abnormal p53 expression, or nuclear localization of ß-Catenin.

Evaluation of quality of life and impact of personality in Chinese obese patients following laparoscopic sleeve gastrectomy.

BACKGROUND/AIMS: The effectiveness of laparoscopic sleeve gastrectomy (LSG) in measuring the quality of life (QOL) and personality in obese patients has not been previously investigated. This study aims to investigate different clinical outcome variables, the relationships between quality of life and different personality one-year after LSG in obese patients. METHODOLOGY: Sixty-one consecutive obese patients (49 females) who underwent LSG were evaluated for clinical characteristics after weight reduction. Chinese Personality Assessment Inventory and Gastrointestinal Quality-of-Life Index (GIQLI) were analyzed. RESULTS: One year after LSG, mean body weight (BW), mean body mass index (BMI) and excess weight loss (EWL) were 66.5±13.6kg, 24.4±4.4kg/m2 and 88.1±32.5%, respectively. Metabolic syndrome decreased from 45.1% to 9.8%. The significant variables were systolic blood pressure, fasting glucose, triglyceride, uric acid, aspartate aminotransferase, alanine aminotransferase, insulin, high sensitivity C-reactive protein, HbA1c, C-peptide and HOMA. All patients demonstrated improvement in two domains of the questionnaire (physical status and emotion status), but not in gastrointestinal symptoms. Although the 'Agreeableness' dimension of Chinese personality had a lower presenting BMI (23.9kg/m2) and better excess weight loss (88.0%), this group showed less improvement in gastrointestinal symptoms and had poorer physical status. CONCLUSIONS: LSG provided meaningful weight loss and improvement in quality of life. Chinese personality was the predictor of weight loss and GIQLI.

Rapid staining and immediate interpretation of fine-needle aspiration cytology for palpable breast lesions: diagnostic accuracy, mammographic, ultrasonographic and histopathologic correlations.

OBJECTIVE: We investigated the role and turn around time of rapid staining and immediate interpretation of fine-needle aspiration cytology (FNAC) for women with palpable breast lesions. STUDY DESIGN: A total of 408 FNAC specimens from 400 patients with palpable breast lesions was analyzed for immediate interpretation and preliminary cytologic diagnosis. All cytological diagnoses were correlated with subsequent alcohol-fixed Papanicolaou-stained slides, mammographic, ultrasonographic and histopathological findings. RESULTS: Of the 408 specimens, 243 (59.6%) were interpreted as benign, 37 (9.0%) atypical, 22 (5.4%) suspicious, 68 (16.7%) malignant, and 38 (9.3%) unsatisfactory. 132 of 408 (32.4%) had subsequent surgical procedures; the sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were 88.5, 100, 100, 81.9 and 92.4%, respectively. The average turn around time was 8.6 min. Mammographic results were available in 242 (59.3%) cases, with 112 (46.3%) undergoing surgical excision. In correlation with mammography and surgical pathology, the false-positive rate, false-negative rate, sensitivity, specificity and accuracy were 1.9, 10.5, 98.1, 89.5 and 95.8%, respectively. CONCLUSIONS: Rapid FNAC interpretation is a useful, effective diagnostic method for palpable breast lesions in our healthcare environment.

Nerve growth factor interacts with CHRM4 and promotes neuroendocrine differentiation of prostate cancer and castration resistance.

Nerve growth factor (NGF) contributes to the progression of malignancy. However, the functional role and regulatory mechanisms of NGF in the development of neuroendocrine prostate cancer (NEPC) are unclear. Here, we show that an androgen-deprivation therapy (ADT)-stimulated transcription factor, ZBTB46, upregulated NGF via ZBTB46 mediated-transcriptional activation of NGF. NGF regulates NEPC differentiation by physically interacting with a G-protein-coupled receptor, cholinergic receptor muscarinic 4 (CHRM4), after ADT. Pharmacologic NGF blockade and NGF knockdown markedly inhibited CHRM4-mediated NEPC differentiation and AKT-MYCN signaling activation. CHRM4 stimulation was associated with ADT resistance and was significantly correlated with increased NGF in high-grade and small-cell neuroendocrine prostate cancer (SCNC) patient samples. Our results reveal a role of the NGF in the development of NEPC that is linked to ZBTB46 upregulation and CHRM4 accumulation. Our study provides evidence that the NGF-CHRM4 axis has potential to be considered as a therapeutic target to impair NEPC progression.

Complete remission of heavily treated ovarian clear cell carcinoma with ARID1A mutations after pembrolizumab and bevacizumab combination therapy: a case report.

BACKGROUND: Patients with ovarian clear cell carcinoma (OCCC) have a poor prognosis because they show low sensitivity to platinum-based chemotherapy. New treatments for refractory OCCC are urgently needed. CASE PRESENTATION: We present a patient with refractory OCCC in whom conventional chemotherapy failed. Cachexia was induced by the disseminating recurrent tumors. Tumor tissue staining and genomic analysis revealed PD-L1 negativity, a low tumor burden, stable microsatellite instability, and two mutations in ARID1A. The patient was administered pembrolizumab combined with bevacizumab triweekly. Her serum CA-125 level decreased dramatically after the first cycle. A computerized tomography scan showed marked regression of the recurrent masses after 3 cycles, and the patient reached complete remission after 9 cycles. She showed good recovery from cachexia. We observed no marked side effects except for mild polyarthritis of the small joints. CONCLUSIONS: The therapeutic effect of checkpoint inhibitors combined with angiogenesis inhibitors is very promising in our patient with OCCC. Further clinical trials of tumors including ARID1A mutations are warranted.

Hypermethylation and decreased expression of TMEM240 are potential early-onset biomarkers for colorectal cancer detection, poor prognosis, and early recurrence prediction.

BACKGROUND: Gene silencing by aberrant DNA methylation of promoter regions remains the most dominant phenomenon occurring during tumorigenesis. Improving the early diagnosis, prognosis, and recurrence prediction of colorectal cancer using noninvasive aberrant DNA methylation biomarkers has encouraging potential. The aim of this study is to characterize the DNA methylation of the promoter region of TMEM240, as well as gene expression and its effect on cell biological functions and its applications in early detection and outcome prediction. RESULTS: Highly methylated CpG sites were identified in the TMEM240 gene by Illumina methylation 450K arrays in 26 Taiwanese patient paired samples and 38 paired samples from The Cancer Genome Atlas (TCGA) colorectal cancer dataset. Transient transfection and knockdown of TMEM240 were performed to demonstrate the role of TMEM240 in colorectal cancer cells. The data showed that TMEM240 could lead to G1 cell cycle arrest, repress cancer cell proliferation, and inhibit cancer cell migration. The quantitative methylation-specific real-time polymerase chain reaction (PCR) results revealed that 87.8% (480 of 547) of the colorectal cancer tumors had hypermethylated TMEM240, and this was also found in benign tubular adenomas (55.6%). Circulating cell-free methylated TMEM240 was detected in 13 of 25 (52.0%) Taiwanese colorectal cancer patients but in fewer (28.6%) healthy controls. In 72.0% (85/118) of tissue samples, TMEM240 mRNA expression was lower in Taiwanese CRC tumor tissues than in normal colorectal tissues according to real-time reverse transcription PCR results, and this was also found in benign tubular adenomas (44.4%). The TMEM240 protein was analyzed in South Korean and Chinese CRC patient samples using immunohistochemistry. The results exhibited low protein expression in 91.7% (100/109) of tumors and 75.0% (24/32) of metastatic tumors but exhibited high expression in 75.0% (6/8) of normal colon tissues. Multivariate Cox proportional hazards regression analysis found that mRNA expression of TMEM240 was significantly associated with overall, cancer-specific, and recurrence-free survival (p = 0.012, 0.007, and 0.022, respectively). CONCLUSIONS: Alterations in TMEM240 are commonly found in Western and Asian populations and can potentially be used for early prediction and as poor prognosis and early-recurrence biomarkers in colorectal cancer.

Prediction of successful weight reduction after laparoscopic adjustable gastric banding.

BACKGROUND/AIM: Compared with conventional pharmacological therapies, bariatric surgery has been shown to cause greater and- sustained weight loss. It was aimed to evaluate weight loss in obese patients after laparoscopic adjustable gastric banding surgery using information typically available during the initial evaluation studied before bariatric surgery and genes. METHODOLOGY: 74 patients undergoing laparoscopic adjustable gastric banding (LAGB) were enrolled. Artificial Neural Network technology was used to predict weight loss. RESULTS: We studied 74 patients consisting of 22 men and 52 women 2 years after operation. Mean age was 31.7 +/- 9.1 years. 27 (36.5%) patients had successful weight reduction (excess weight loss >50%) while 47 (63.5%) did not. ANN provided predicted factors on gender, insulin, albumin and two genes: re4684846_r, rs660339_r which were associated with success. CONCLUSION: Artificial neural network is a better modeling technique and the predictive accuracy is higher on the basis of multiple variables related to laboratory tests. Our finding gave demonstrated result that obese patients of successful weight reduction after laparoscopic adjustable gastric banding surgery were women, having little lower insulin and albumin, and carrying GG genotype on rs4684846 and with at least one T allele on rs660339. In these cases, weight loss will give better results.

Obesity and the decision tree: predictors of sustained weight loss after bariatric surgery.

BACKGROUND/AIMS: Bariatric surgery is the only long-lasting effective treatment to reduce body weight in morbid obesity. Previous literature in using data mining techniques to predict weight loss in obese patients who have undergone bariatric surgery is limited. This study used initial evaluations before bariatric surgery and data mining techniques to predict weight outcomes in morbidly obese patients seeking surgical treatment. METHODOLOGY: 251 morbidly obese patients undergoing laparoscopic mini-gastric bypass (LMGB) or adjustable gastric banding (LAGB) with complete clinical data at baseline and at two years were enrolled for analysis. Decision Tree, Logistic Regression and Discriminant analysis technologies were used to predict weight loss. Overall classification capability of the designed diagnostic models was evaluated by the misclassification costs. RESULTS: Two hundred fifty-one patients consisting of 68 men and 183 women was studied; with mean age 33 years. Mean +/- SD weight loss at 2 year was 74.5 +/- 16.4 kg. During two years of follow up, two-hundred and five (81.7%) patients had successful weight reduction while 46 (18.3%) were failed to reduce body weight. Operation methods, alanine transaminase (ALT), aspartate transaminase (AST), white blood cell counts (WBC), insulin and hemoglobin A1c (HbA1c) levels were the predictive factors for successful weight reduction. CONCLUSION: Decision tree model was a better classification models than traditional logistic regression and discriminant analysis in view of predictive accuracies.