SEPTIN9-SDC2-VIM methylation signature as a biomarker for the early diagnosis of colorectal cancer.

The accurate detection of colorectal cancer (CRC) at its initial stage can reduce mortality. However, the broad application of endoscopy has been limited due to the invasive procedure and patient noncompliance. Liquid biopsy with subsequent mapping of methylation in specific cell-free DNA (cfDNA) may represent an alternative approach for early diagnosis. In this study, we have developed a minimal-invasive blood-based test for detection of precancerous lesions and early-stage CRC. Using TCGA M450K methylation data, we identified candidate methylation sites with the highest Fold Change (FC) for three genes (SEPTIN9, SDC2 and VIM), which were selected from previous studies. Based on logistic regression models, we developed a 3-gene methylation signature for CRC diagnosis with high accuracy (Sensitivity =0.959, Specificity =1, AUC =0.997). Using independent public databases and data from blood samples, this model has demonstrated superior performance. The AUC was 0.919-1 and 0.905-0.916 in public tissue database for CRC and blood sample data, respectively. Thus, our proposed 3-gene methylation signature has a more reliable performance than other methods. Furthermore, signal enhancement effect of 3-gene methylation signature can improve the accuracy of early diagnosis for CRC, which demonstrates the potential for clinical application.

Response of Soil Nematode Community Structure and Function to Monocultures of Pumpkin and Melon.

It is well known that crop monoculture can induce negative effects on soil ecosystems and crop productivity. However, little is known about how vegetable monoculture affects the soil nematode community structure and its relationship with vegetable yields. In this study, the composition, abundance, metabolic footprint, and ecological indices of soil nematodes are investigated in monocultures of pumpkin and melon. The relationships between nematode community structure and yields of pumpkin and melon were analyzed by linear regression. Both monoculture soils of pumpkin and melon suppressed the relative abundance of bacterivores but increased the relative abundance of plant parasites. Pumpkin monoculture soils decreased soil nematode diversity but increased the maturity index of plant parasites. Monoculture soils of pumpkin and melon decreased the metabolic footprint of lower- and higher-level trophic groups of the soil food web, respectively. Pumpkin and melon monoculture soils increased the food web indices channel index (CI) but decreased the enrichment index (EI) and the structure index (SI). The monoculture soils of pumpkin and melon led to a more fungal-dominated decomposition pathway and degraded soil food web conditions. The abundance of bacterivores and food web indices EI and SI were positively correlated with soil nutrients and pH, while the abundance of plant parasites and CI were negatively correlated with soil nutrients and pH. Paratylenchus was negatively correlated with pumpkin and melon yields and could be the potential plant parasites threatening pumpkin and melon productions. Redundancy analysis showed that monocultures of pumpkin and melon altered the soil nematode community via soil properties; total N, total P, alkeline-N, and pH were the main driving factors.

Risk and Prognosis of Subsequent Primary Gastric Cancer.

INTRODUCTION: The incidence and prognostic impact of subsequent primary gastric cancer (GC) in a population of other cancer survivors is unclear. We aimed to evaluate susceptibility to subsequent primary GC in cancer survivors and prognosis of GC with prior cancer history. METHODS: A total of 2,211 and 23,416 GC cases with and without prior cancer history were retrospectively selected from the Surveillance, Epidemiology, and End Results (SEER) database. Potential risk of developing subsequent primary GC was assessed through standardized incidence ratios (SIRs). Cox regression was adopted to analyze the influence of prior cancer history and clinical characteristic factors on the prognosis of subsequent primary GC. A nomogram was established to predict overall survival (OS). Propensity score matching was conducted to eliminate possible bias. RESULTS: Compared with general population, cancer survivors had an increased risk of subsequent primary GC (SIR 1.17, 95% CI: 1.15-1.20, p < 0.05). Prior cancer history was related to poor OS of GC (adjusted hazard ratio [aHR] 1.12, 95% CI: 1.06-1.19, p < 0.001), but not cancer-specific survival (aHR 0.97, 95% CI: 0.89-1.05, p = 0.441). In addition, age, grade, stage, year of diagnosis, surgery, TNM stage, and tumor size were independent prognostic factors for OS in GC cases with prior cancers. The concordance index of the nomogram was 0.72 (95% CI: 0.71-0.74), and calibrate curves showed good agreement between prediction by the nomogram and actual observation. CONCLUSIONS: Cancer survivors with increased risk of developing subsequent primary GC should strengthen their monitoring and follow-up to prevent occurrence of subsequent primary GC.

ZRANB1 enhances stem-cell-like features and accelerates tumor progression by regulating Sox9-mediated USP22/Wnt/ß-catenin pathway in colorectal cancer.

The pathogenesis of colorectal cancer (CRC) is a multistep process characterized by the accumulation of gene mutations and epigenetic alterations. Tumor necrosis factor receptor-associated factor-binding protein domain (ZRANB1) is a deubiquitinase that mediates tumor growth and metastasis by deubiquitinating target proteins. In this study, we examined the regulatory effects of ZRANB1 on the maintenance of cancer stem cell (CSC) properties and tumor growth in CRC. Human CRC tissue samples and matched normal tissues were collected for the analysis of ZRANB1 expression. ZRANB1 was upregulated in CRC human tissues and cell lines, and its expression was positively correlated with advanced tumor stage and poor survival of CRC patients. The overexpression of ZRANB1 also induced the expression of CSC markers in CRC cells. Then, a xenograft model was established by inoculating BALB/c mice with CRC cells. The upregulation of ZRANB1 promoted tumorigenesis in vivo. Sox9 is a transcription factor that acts as an oncogene in human cancers. ZRANB1 increased the stability of Sox9 in CRC cells by decelerating its ubiquitination. Further analysis revealed that Sox9 regulated the transcription activity of USP22 by binding to its promoter. Moreover, ZRANB1 enhances stem-cell-like features of CRC cells and activated the Wnt/ß-catenin pathway through USP22. Our results highlighted the role of ZRANB1 as a molecular target for CRC treatment, which may contribute to the development of novel therapies with better efficacy.

Nonreceptor tyrosine phosphatase 14 promotes proliferation and migration through regulating phosphorylation of YAP of Hippo signaling pathway in gastric cancer cells.

BACKGROUND: The Hippo signaling pathway is associated with cell proliferation and organ size, and its transcriptional coactivator Yes-associated protein (YAP), emerges as a crucial oncoprotein in multiple cancers. It was increasingly recognized that nonreceptor tyrosine phosphatase 14 (PTPN14) was relevant to the cell membrane and cytoskeleton, and had a critical effect on cell adhesion, growth, and actin cytoskeleton organization. Furthermore, PTPN14 was also certified to operate the translocation and phosphorylation of YAP. The present experiment was aimed to explore the impact of PTPN14 on gastric cancer (GC) cell proliferation and migration through regulating the phosphorylation of YAP. METHODS: The pEGFP-N1-PTPN14 recombinant plasmid was stably transfected into three differentiation degrees GC cell lines, including MKN-28, SGC-7901, and BGC-823. Quantitative reverse transcription-polymerase chain reaction and Western blot assay were performed to analyze the messenger RNA (mRNA) and protein levels. The proliferative and migratory capacity of cells was appraised by Cell Counting Kit-8 assay and transwell chamber. RESULTS: Compared with the normal control and vector transfection group, the capacity of these three cell lines, which transfected with the pEGFP-N1-PTPN14 to proliferate and migrate in vitro was increased obviously (P < .05). There was no YAP mRNA detected in MKN-28 cell line. Meanwhile, after transfecting the pEGFP-N1-PTPN14 plasmid, the mRNA level of YAP in SGC-7901 was reduced (P < .05), and it was increased in BGC-823 (P < .05). The YAP protein level in SGC-7901 and BGC-823 has no apparent transformation by transfecting, but the protein level of phospho-Ser127 YAP and phospho-Ser397 YAP is upregulated (P < .05). CONCLUSION: PTPN14 could enhance the proliferative and migratory ability of GC cells by promoting the YAP phosphorylation in the Hippo signaling pathway. Taken together, PTPN14 might be involved in the occurrence and development of GC and become a molecular regulator to treat GC.

Aberrant methylation of FAT4 and SOX11 in peripheral blood leukocytes and their association with gastric cancer risk.

Background: Aberrant DNA methylation, especially tumor suppressor gene hypermethylation, is a well-recognized biomarker of initial tumorogenesis stages. FAT4 and SOX11 are putative tumor suppressor genes and can be down-regulated by hypermethylation in various cancers tissues. However, in peripheral blood leukocytes, the association between these two genes methylation status, as well as the effects of gene-environment interactions, and gastric cancer (GC) risk remain unclear. Methods: A hospital-based case-control study including 375 cases and 394 controls was conducted. Peripheral blood leukocytes DNA methylation status were detected by methylation-sensitive high-resolution melting (MS-HRM) assay. Logistic regression was adopted to analyze the relationship of FAT4 and SOX11 methylation with GC susceptibility. Results: Positive methylation (Pm) and total positive methylation (Tpm) of FAT4 were significantly increased the risk of GC (OR = 2.204, 95% CI: 1.168-4.159, P = 0.015; OR = 1.583, 95% CI: 1.031-2.430, P = 0.036, respectively). Compared with controls, cases exhibited higher SOX11 Pm frequencies with OR of 2.530 (95% CI: 1.289-4.969, P = 0.007). Nonetheless, no statistically significant association between SOX11 Tpm and GC risk was observed. Additionally, interactions between FAT4 Tpm and increased consumption of freshwater fish (≥1 times/week) displayed an antagonistic effect on GC (OR = 0.328, 95% CI: 0.142-0.762, P = 0.009), and high salt intake interacted with SOX11 Tpm also showed statistically significant (OR = 0.490, 95% CI: 0.242-0.995, P = 0.048). Conclusions:FAT4 aberrant methylation in peripheral blood leukocytes and gene-environment interactions were associated with the risk of GC, while SOX11 was controversial and needed to be more investigated.

The lncRNA CRNDE promotes colorectal cancer cell proliferation and chemoresistance via miR-181a-5p-mediated regulation of Wnt/ß-catenin signaling.

BACKGROUND: With more than 600,000 mortalities each year, colorectal cancer (CRC) is the third most commonly diagnosed type of cancer worldwide. Recently, mechanisms involving noncoding RNAs have been implicated in the development of CRC. METHODS: We examined expression levels of lncRNA CRNDE and miR-181a-5p in 64 cases of CRC tissues and cell lines by qRT-PCR. Gain-of-function and loss-of-function assays were performed to examine the effect of CRNDE and miR-181a-5p on proliferation and chemoresistance of CRC cells. Using fluorescence reporter and western blot assays, we also explored the possible mechanisms of CRNDE in CRC cells. RESULTS: In this study, we found that the expression levels of the CRNDE were upregulated in CRC clinical tissue samples. We identified microRNA miR-181a-5p as an inhibitory target of CRNDE. Both CRNDE knockdown and miR-181a-5p overexpression in CRC cell lines led to inhibited cell proliferation and reduced chemoresistance. We also determined that ß-catenin and TCF4 were inhibitory targets of miR-181a-5p, and that Wnt/ß-catenin signaling was inhibited by both CRNDE knockdown and miR-181a-5p overexpression. Significantly, we found that the repression of cell proliferation, the reduction of chemoresistance, and the inhibition of Wnt/ß-catenin signaling induced by CRNDE knockdown would require the increased expression of miR-181a-5p. CONCLUSIONS: Our study demonstrated that the lncRNA CRNDE could regulate the progression and chemoresistance of CRC via modulating the expression levels of miR-181a-5p and the activity of Wnt/ß-catenin signaling.

rs744166 polymorphism of the STAT3 gene is associated with risk of gastric cancer in a Chinese population.

The aim of this study was to explore the association between polymorphisms in signal transducer and activator of transcription protein 3 (STAT3) and the risk of gastric cancer. In the present study, a case-control study was conducted in which rs2293152 and rs744166 polymorphisms in STAT3 were analyzed in 209 Chinese patients with gastric cancer and 294 cancer-free controls. The genotypes were determined by polymerase chain reaction restriction fragment length polymorphism method. For the rs744166 polymorphism, the TC genotype (adjusted OR = 0.60, 95% CI = 0.39-0.92, and P = 0.020) and CC genotype (adjusted OR = 0.41, 95% CI = 0.21-0.80, and P = 0.009) were associated with a decreased risk of gastric cancer compared to the TT genotype. However, rs2293152 did not show any difference in gastric cancer risk between patients and controls in the CG/CC genotype compared to the GG genotype. Besides, the SNP effects were additive to the effects of environmental factors without any interaction between them in the susceptibility to gastric cancer. Collectively, rs744166 polymorphism might be significantly associated with a decreased risk of gastric cancer in a Chinese population. Additionally, polymorphisms in STAT3, along with environmental factors, might be associated with the development of gastric cancer.

Serum Helicobacter pylori NapA antibody as a potential biomarker for gastric cancer.

Helicobacter pylori (H. pylori) infection is strongly associated with gastric cancer. However, only a minority of infected individuals ever develop gastric cancer. This risk stratification may be in part due to differences among strains. The relationship between neutrophil-activating protein (NapA) and gastric cancer is unclear. The purpose of this study is to evaluate the significance of NapA as a biomarker in gastric cancer. We used enzyme linked immunosorbent assay (ELISA) to determine the status of H. pylori infection. Indirect ELISA method was used for detection of NapA antibody titer in the serum of H. pylori infected individuals. Unconditional logistic regressions were adopted to analyze the variables and determine the association of NapA and gastric cancer. The results of study indicated serum H. pylori NapA antibody level were associated with a reduced risk for development of gastric cancer. It may be used in conjugation with other indicators for gastric cancer detection.

Serum antibody against Helicobacter pylori FlaA and risk of gastric cancer.

BACKGROUND: Helicobacter pylori (H. pylori) infection is a major risk factor for gastric cancer (GC); however, only a minority of infected individuals develops GC. We aim to assess the association between serostatus of antibody against H. pylori flagellin A (FlaA) and risk of GC and to evaluate the value of serum FlaA antibody as a novel screening biomarker for GC risk. METHODS: A hospital-based case-control study including 232 cases and 264 controls was conducted. Logistic regression was adopted to analyze the association between the serostatus of FlaA antibody and risk of GC. Serum FlaA antibody was measured by an enzyme-linked immunosorbent assay (ELISA). Receiver operating characteristic (ROC) curve was used to evaluate the screening efficacy and to identify a cutoff point of serum FlaA antibody level. RESULTS: Helicobacter pylori infection was associated with an increased risk of GC (p = .007). A positive association between serum FlaA antibody and GC risk was observed in overall subjects and H. pylori-positive subjects (OR [95% CI]: 6.8 [4.3-10.7] and 6.9 [3.6-13.4], respectively; p < .001). The seropositivity of FlaA antibody was strongly related to GC risk in a dose-dependent manner (p for trend < .001). The optimal cutoff value (OD) was 0.1403, providing a sensitivity of 74.1% and a specificity of 64.4%. The area under the ROC curve (AUC) was 0.74 in overall subjects and 0.73 in H. pylori-positive subjects, respectively. CONCLUSIONS: FlaA was an independent risk factor for H. pylori-related GC. Serum FlaA antibody may serve as a novel noninvasive biomarker for early detection of GC.

Polymorphisms of the TLR4 gene and risk of gastric cancer.

OBJECTIVE: Toll-like receptor 4 (TLR4) is an important lipo-polysaccharide (LPS) receptor in gastric epithelial cell signaling transduction and plays critical roles in the development and progression of gastric cancer (GC). We investigated the effects of TLR4 gene polymorphisms and gene-environmental interactions on the risk of GC in Northeastern China. METHODS: We genotyped two single-nucleotide polymorphisms (SNPs) in TLR4 (rs10116253 and rs1927911) in 217 GC patients and 294 cancer-free controls using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Odds ratio (OR) and 95% confidence intervals (CIs) were estimated by unconditional logistic-regression models. RESULTS: Individuals carrying CC genotype of rs10116253 and TT genotype of rs1927911 had a significantly decreased risk of GC (adjusted OR=0.33, 95% CI 0.18-0.60, P<0.001 and adjusted OR=0.37, 95% CI 0.21-0.67, P=0.001 respectively), compared with TT genotype of rs10116253 and CC genotype of rs1927911. In addition, the SNP effects were additive to the effects of some known environmental factors without any interaction between them in the susceptibility to GC. CONCLUSION: Our data suggested that TLR4 gene polymorphisms may be associated with a decreased risk of GC in Chinese population. And these SNPs and their combined effects with environmental factors may be associated with the risk of GC.

The influence of genetic polymorphisms in MDR1 gene on breast cancer risk factors in Chinese.

Breast cancer (BC) is the most common cancer among women in the world. The human multidrug resistance 1 gene (MDR1) is potentially an important gene influencing the susceptibility to breast cancer. This study aimed to assess the association of MDR1 genetic polymorphisms with the susceptibility to BC. Overall, 353 BC patients and 360 cancer-free controls were enrolled. The clinical characteristics were summarized by questionnaires. The c.1564A > T genetic polymorphism was genotyped using created restriction site-polymerase chain reaction method. We found that no significant differences in the genotypic and allelic frequencies between BC patients and cancer-free controls. Furthermore, the distribution of BC patients' risk factors was not significantly different among AA, AT, and TT genotypes. Our findings indicate that the c.1564A > T genetic polymorphism is not significantly associated with the susceptibility to BC in Chinese Han populations.

Tumor necrosis factor signaling is implicated in intestinal mucosal injury of cancer cachexic mice.

This study will examine the relationship between tumor necrosis factor (TNF) and intestinal mucosal injury in a cancer cachexia mouse model. The C26 adenocarcinoma cancer model was set up, and immunohistochemistry was performed to investigate the histological chances of injuring the intestinal mucosa and the intestinal villi. The liquid-phase protein chip was employed to examine the changes in TNF and its receptors. The results demonstrated an intestinal mucosal injury in the cancer cachexia mouse model. The height of the intestinal villi decreased, and thinner basal membrane thickness was noted. The serum TNF-alpha and sTNFR1 increased, while in the mucosa, the TNF-alpha concentration increased; sTNFR1 and sTNFR2 decreased when compared to the control group. In conclusion, there is a potential association between the TNF signaling pathway and the intestinal mucosal injury in a cancer cachexia mouse model. Such understanding provides insights into the development of novel therapeutic targets for intestinal mucosa protection in clinical practice.

Pathway-specific analysis of gene expression data identifies the PI3K/Akt pathway as a novel therapeutic target in cervical cancer.

PURPOSE: Cervical tumor response on posttherapy 2[(18)F]fluoro-2-deoxy-d-glucose-positron emission tomography (FDG-PET) is predictive of survival outcome. The purpose of this study was to use gene expression profiling to identify pathways associated with tumor metabolic response. EXPERIMENTAL DESIGN: This was a prospective tissue collection study for gene expression profiling of 62 pretreatment biopsies from patients with advanced cervical cancer. Patients were treated with definitive radiation. Fifty-three patients received concurrent chemotherapy. All patients underwent a pretreatment and a 3-month posttherapy FDG-PET/computed tomography (CT). Tumor RNA was harvested from fresh frozen tissue and hybridized to Affymetrix U133Plus2 GeneChips. Gene set enrichment analysis (GSEA) was used to identify signaling pathways associated with tumor metabolic response. Immunohistochemistry and in vitro FDG uptake assays were used to confirm our results. RESULTS: There were 40 biopsies from patients with a complete metabolic response (PET-negative group) and 22 biopsies from patients with incomplete metabolic response (PET-positive group). The 3-year cause-specific survival estimates were 98% for the PET-negative group and 39% for the PET-positive group (P < 0.0001). GSEA identified alterations in expression of genes associated with the PI3K/Akt signaling pathway in patients with a positive follow-up PET. Immunohistochemistry using a tissue microarray of 174 pretreatment biopsies confirmed p-Akt as a biomarker for poor prognosis in cervical cancer. The phosphoinositide 3-kinase (PI3K) inhibitor LY294002 inhibited FDG uptake in vitro in cervical cancer cell lines. CONCLUSIONS: Activation of the PI3K/Akt pathway is associated with incomplete metabolic response in cervical cancer. Targeted inhibition of PI3K/Akt may improve response to chemoradiation.

[Clinical experience of percutaneous endoscopic gastrostomy, jejunostomy, duodenostomy in 120 patients].

OBJECTIVE: To report clinical experience of percutaneous endoscopic gastrostomy, duodenostomy, jejunostomy in 120 patients, focusing on its technique and indications. METHODS: One hundred and twenty patients received percutaneous endoscopic gastrostomy, duodenostomy, jejunostomy from May 2001 to April 2004, including 75 percutaneous endoscopic gastrostomy (PEG), 42 percutaneous endoscopic jejunostomy (PEJ), 2 percutaneous endoscopic duodenostomy (PED), 1 direct percutaneous endoscopic jejunostomy (DPEJ). All tubes established by traditional pull technique. RESULTS: The average duration of PEG was (9 +/- 4) min, PEJ (17 +/- 6) min, DPEJ 20 min, and PED was 10 and 12 min for 2 patients, respectively. Success rate of the technique was 98.4% (120/122). Major complication rate was 0.8% (1/120), and minor complication rate was 7.5% (9/120). Clinical indications: PEG, PED and PEJ were applied for long-term enteral nutritional support in 88 patients, gastrointestinal decompression in 25 patients, and transfusing external drainage bile to gastrointestinal tract in 5 patients. Two radiation enteritis patients used PEG for gastrointestinal decompression preoperatively and long-term enteral nutritional support postoperatively. CONCLUSION: PEG, PED PEJ and DPEJ are easily handled, effective and safe, and may be widely used in clinical practice.

Effects of endostatin on expression of vascular endothelial growth factor and its receptors and neovascularization in colonic carcinoma implanted in nude mice.

AIM: To investigate the antiangiogenic effects of endostatin on colonic carcinoma cell line implanted in nude mice and its mechanism. METHODS: Nude mice underwent subcutaneous injection with LS-174t colonic carcinoma cell line to generate carcinoma and were randomly separated into two groups. Mice received injection of vehicle or endostatin every day for two weeks. After the tumor was harvested, the tumor volumes were determined, and the expressions of CD34, VEGF and Flk-1 were examined by immunohistochemical method. RESULTS: Tumor volume was significantly inhibited in the endostatin group (84.17%) and tumor weight was significantly inhibited in the endostatin group (0.197+/-0.049) compared to the control group (1.198+/-0.105) (F = 22.56, P = 0.001), microvessel density (MVD) was significantly decreased in the treated group (31.857+/-3.515) compared to the control group (100.143+/-4.290) (F = 151.62, P<0.001). Furthermore, the expression of Flk-1 was significantly inhibited in the treated group (34.29%) compared to the control group (8.57%) (chi(2) = 13.745, P = 0.001). However no significant decrease was observed in the expression of vascular endothelial growth factor (VEGF) between these two groups (chi(2) = 0.119,P = 0.730). CONCLUSION: Endostatin can inhibit tumor growth and angiogenesis by blocking Vegf/Flk-1 pathway. This experiment provides the theory basis for developing a new anti-carcinoma drug through studying the properties of anti-angiogenesis inhibitors.