Normal Stomach Research Articles

Laparoscopic ultrasound guided wedge resection of the stomach: a novel procedure for gastric submucosal tumor.

It is essential for the surgery of gastric submucosal tumors to resect the tumor with a negative margin and minimize the incision of the normal stomach wall. We developed a novel procedure for patients with gastric submucosal tumors using a laparoscopic ultrasound probe as a guide to determine the resection line. Since 2014, we have performed the laparoscopic ultrasound-guided wedge resection of the stomach in seven patients. The tumor was localized, and the property of the tumor was clearly identified using a laparoscopic ultrasound probe. As a result, the ideal incision line was determined without intraoperative endoscopy. The stomach wall was perforated along the marking on the planned incision line and the whole layer is subsequently incised along with the tumor. The surgical margins were negative, and there were no obvious injuries of the pseudocapsule, microscopically, in any case. It is possible that the laparoscopic ultrasound-guided wedge resection of the stomach contributes to a simplification of the surgery of gastric submucosal tumors resulting in reduced medical cost while maintaining curability and functional preservation.

The Volatilomic Footprints of Human HGC-27 and CLS-145 Gastric Cancer Cell Lines.

The presence of certain volatile biomarkers in the breath of patients with gastric cancer has been reported by several studies; however, the origin of these compounds remains controversial. In vitro studies, involving gastric cancer cells may address this problem and aid in revealing the biochemical pathways underlying the production and metabolism of gastric cancer volatile indicators. Gas chromatography with mass spectrometric detection, coupled with headspace needle trap extraction as the pre-concentration technique, has been applied to map the volatilomic footprints of human HGC-27 and CLS-145 gastric cancer cell lines and normal Human Stomach Epithelial Cells (HSEC). In total, 27 volatile compounds are found to be associated with metabolism occurring in HGC-27, CLS-145, and HSEC. Amongst these, the headspace concentrations of 12 volatiles were found to be reduced compared to those above just the cultivating medium, namely there was an observed uptake of eight aldehydes (2-methylpropanal, 2-methyl-2-propenal, 2-methylbutanal, 3-methylbutanal, hexanal, heptanal, nonanal, and benzaldehyde), three heterocyclic compounds (2-methyl-furan, 2-ethyl-furan, and 2-pentyl-furan), and one sulfur-containing compound (dimethyl disulphide). For the other 15 volatiles, the headspace concentrations above the healthy and cancerous cells were found to be higher than those found above the cultivating medium, namely the cells were found to release three esters (ethyl acetate, ethyl propanoate, and ethyl 2-methylbutyrate), seven ketones (2-pentanone, 2-heptanone, 2-nonanone, 2-undecanone, 2-tridecanone, 2-pentadecanone, and 2-heptadecanone), three alcohols (2-methyl-1-butanol, 3-methyl-1-butanol, and 2-ethyl-1-hexanol), one aromatic compound (toluene), and one sulfur containing compound [2-methyl-5-(methylthio) furan]. In comparison to HSEC, HGC-27 cancer cell lines were found to have significantly altered metabolism, manifested by an increased production of methyl ketones containing an odd number of carbons. Amongst these species, three volatiles were found exclusively to be produced by this cell line, namely 2-undecanone, 2-tridecanone, and 2-heptadecanone. Another interesting feature of the HGC-27 footprint is the lowered level of alcohols and esters. The CLS-145 cells exhibited less pronounced changes in their volatilomic pattern compared to HSEC. Their footprint was characterized by the upregulated production of esters and 2-ethyl-hexanol and downregulated production of other alcohols. We have therefore demonstrated that it is possible to differentiate between cancerous and healthy gastric cells using biochemical volatile signatures.

Gastritis: Sociodemographic, Clinical, Endoscopic and Histological Aspects, about 593 Cases at the Digestive Endoscopy Unit of the General Hospital Idrissa Pouye

Introduction: Gastritis is a very common and widely distributed condition worldwide. It represents one of the most common pathological entities in gastroenterology and digestive endoscopy. Our objective was to determine the sociodemographic, clinical, endoscopic, and histological aspects of gastritis in the digestive endoscopy unit of the General Hospital Idrissa Pouye (GHIP). Materials and Method: This was a retrospective study over a period of 4 years (from 1 January 2014 to 31 December 2017) at the digestive endoscopy unit of GHIP. We had collated oesogastroduodenal endoscopy (EGDE) reports with gastritis appearance with gastric biopsies and reports with normal stomach appearance with gastric biopsies and their histological reports. We collected and analyzed data on age, gender, indications for endoscopy, endoscopic findings and histological results. Results: The reports of 593 patients were analyzed. The mean age was 45 years old (range 8 - 88 years old) and the sex ratio was 0.63 (230 men). The indications for endoscopy were epigastralgia in (91%) of cases, dyspepsia in (22%) of cases, pyrosis in (12%) of cases. The endoscopic appearance was normal in 229 patients (39%). The endoscopic location of the gastritis was antral in 76%, fundic in 22% and pangastric in 2%. The gastritis was erythematous in 327 patients (90%), erosive in 126 patients (35%), congestive in 53 patients (15%), pseudonodular in 14 patients (4%) and atrophic in 10 patients (3%). Histology was normal in 8 patients (1.3%) and showed gastritis in 585 patients (98.7%). Gastritis was chronic in 575 patients (98.2%), acute in 10 patients (1.7%). Gastritis activity was moderate in (52.7%) and mild in (42.9%). Atrophy was absent in 521 patients (88.6%) and mild in 46 patients (8.2%). Intestinal metaplasia was found in 66 patients (11%). Dysplasia was present in 1.7% of cases. This dysplasia was intermediate grade (60%) in 6 patients, low grade (20%) in 2 patients and severe grade (20%). H. pylori was present in 404 patients (68%). Conclusion: Gastritis is usually found in the digestive endoscopy unit of the GHIP. The indications for endoscopy are dominated by epigastralgia and histology is necessary for its diagnosis.

GINS Complex Subunit 2 Facilitates Gastric Adenocarcinoma Proliferation and Indicates Poor Prognosis.

Gastric cancer is the one of the most lethal malignancies of digestive system. Identifying molecular biomarkers is invaluable in help predicting clinical outcomes and developing targeted chemotherapies. GINS complex subunit 2 (GINS2) plays an essential role in the initiation and elongation of DNA replication. Although there have been studies revealing the prognostic significance of GINS2 in breast cancer and lung cancer, its involvement and function in gastric cancer need to be elucidated. We retrospectively enrolled a cohort of gastric adenocarcinoma patients after surgical resection (n = 123). By analyzing the mRNA and protein levels of GINS2 in tissue samples, we found that GINS2 presented a higher expression in tumor tissues than in adjacent normal stomach tissues. Besides, GINS2 level was positively correlated with tumor size and gastric adenocarcinoma tumor stage, implying its potential role as a tumor promoter. Univariate and multivariate analyses identified that patients with lower GINS2 showed a better overall survival compared to those with higher GINS2 expression. In addition, cellular and xenograft experiments confirmed the role of GINS2 in facilitating tumor proliferation both in vitro and in vivo. To our knowledge, this is the initial finding on GINS2 in promoting gastric adenocarcinoma progression. In conclusion, our study revealed a pro-oncogenic role of GINS2 in gastric cancer.

Gastric peroral myotomy (G-POEM) in patients with delayed emptying function of the stomach

Introduction. Disorders in the gastric evacuation function are met in patients with gastroesophageal reflux. In most cases, simultaneous laparoscopic pyloroplasty can solve the problem. As an alternative to surgical pyloroplasty to treat gastroparesis, a new technique has been proposed – peroral submucosal pyloromyotomy or gastric peroral endoscopic myotomy (G-POEM). Recently, this endoscopic surgery has been implemented in adults and in newborns with congenital hypertrophic pyloric stenosis to treat gastric paresis. In the present work, the authors describe this technique and short-term results after G-POEM in a child who had previously been operated on for gastroesophageal reflux.Material and methods. Peroral submucosal pyloromyotomy was performed in a girl of 5 y.o. with impaired motor-evacuation function of the stomach after surgical treatment of gastroesophageal reflux disease at the age of two . For 3 years, the child had therapy which included antiemetics and H2-receptor blockers. However, clinical symptoms constantly recurred, and it was decided to make a thorough examination of the patient. At the contrast examination of upper gastrointestinal tract, stomach enlargement and slow transit of the contrast preparation into the duodenum were seen. Fibrogastroduodenoscopy confirmed the normal function of the fundoplication cuff and found a cause of impaired gastric emptying. A contracted gastric outlet as a spasmodic pylorus was found to be an obstacle for normal stomach evacuation function. A narrow hole in the pylorus was found. A 5.4 mm gastroscope could pass through it. The patient had a complete peroral endoscopic submucosal pyloromyotomy. The technique of this surgery consisted in creating a submucosal tunnel at 4 cm before the pylorus and in dissecting the hypertrophied muscle layer by Ramstedt incision using an electrocoagulation knife. After the end of surgery, the incision on the mucous layer was closed with special clamps.Results. Surgical time lasted for 45 minutes. There were no intraoperative complications: bleedings and mucosal perforation . The patient began to eat in 6 hours after the surgery. The transition to full enteral feeding lasted for 24 hours. The child was discharged from the hospital next day in good condition. At the follow-up examination 6 months after surgery, the girl had weight and height typical for her age. There were no signs of dysphagia, abdominal distension, nausea and vomiting any more . There were no post-operative scarring on the child’s abdominal wall.Conclusion. Peroral submucosal pyloromyotomy is technically implementable, safe and effective for treating disorders of stomach evacuation function in children. Further research is needed to find the place of this technique in the treatment of gastric outlet obstruction.

Autoimmune Gastritis and Gastric Microbiota.

Autoimmune atrophic gastritis is an organ-specific immune-mediated condition characterized by atrophy of the oxyntic mucosa. Autoimmune atrophic gastritis (AIG) is characterized by a progressive loss of acid-secreting parietal cells leading to hypo-achlorhydria. Due to this peculiar intra-gastric environment, gastric microbiota composition in individuals with autoimmune atrophic gastritis was first supposed and then recently reported to be different from subjects with a normal acidic healthy stomach. Recent data confirm the prominent role of Helicobacter pylori as the main bacterium responsible for gastric disease and long-term complications. However, other bacteria than Helicobacter pylori, for example, Streptococci, were found in subjects who developed gastric cancer and in subjects at risk of this fearful complication, as well as those with autoimmune gastritis. Gastric microbiota composition is challenging to study due to the acidic gastric environment, the difficulty of obtaining representative samples of the entire gastric microbiota, and the possible contamination by oral or throat microorganisms, which can potentially lead to the distortion of the original gastric microbial composition, but innovative molecular approaches based on the analysis of the hyper-variable region of the 16S rRNA gene have been developed, permitting us to obtain an overall microbial composition view of the RNA gene that is present only in prokaryotic cells.

FOXD1-AS1 regulates FOXD1 translation and promotes gastric cancer progression and chemoresistance by activating the PI3K/AKT/mTOR pathway.

Gastric cancer (GC) is a common gastrointestinal cancer with a high global mortality. Recent reports have suggested that long noncoding RNA (lncRNA) are implicated in multiple aspects of GC, including pathogenesis, progression, and therapeutic response. Herein, we investigated the function of FOXD1‐AS1 in GC progression and chemoresistance. Expression of FOXD1‐AS1 was low in normal stomach tissues but was upregulated in GC cell lines. Silencing of FOXD1‐AS1 impaired GC cell proliferation and motility in vitro, and repressed tumor growth and metastasis in vivo. Importantly, FOXD1‐AS1 upregulation increased the resistance of GC cells to cisplatin. Moreover, we found that FOXD1‐AS1 promoted FOXD1 protein translation through the eIF4G‐eIF4E‐eIF4A translational complex. We also demonstrated that FOXD1‐AS1 released eIF4E from phosphorylated 4E‐BP1 and thereby strengthened the interaction of eIF4E with eIF4G by activating the PI3K/AKT/mTOR pathway. Activation of the PI3K/AKT/mTOR pathway was due to the post‐transcriptional upregulation of PIK3CA, in turn induced by FOXD1‐AS1‐mediated sequestering of microRNA (miR)‐466. Furthermore, we verified that FOXD1‐AS1 facilitated GC progression and cisplatin resistance in a FOXD1‐dependent manner. In conclusion, FOXD1‐AS1 aggravates GC progression and chemoresistance by promoting FOXD1 translation via PIK3CA/PI3K/AKT/mTOR signaling. These findings highlight a novel target for treatment of patients GC, particularly patients with cisplatin resistance.

Application of Computed Tomography Imaging in Diagnosis of Endocrine Nerve of Gastric Cancer and Nursing Intervention Effect

In this article, some parameters and characteristics of computed tomography (CT) images in patients with gastric cancer are analyzed and the application of CT images in the diagnosis of gastric cancer endocrine nerves and the impact of nursing intervention on the quality and mental state of CT images of patients with gastric cancer are discussed. First, all patients were scanned with CT, and the CT values of the normal stomach wall and all lesions at different single-energy levels were recorded separately. Second, the improved back propagation network model was applied to realize the diagnosis of gastric cancer through the analysis of various features of CT images. The effect of nursing intervention on the image quality and mental state of CT imaging of patients with gastric cancer was studied. The results show that the energy spectrum curve of CT images and the improved back propagation network model are helpful for the initial diagnosis and identification of gastric cancer. Nursing intervention has a good influence on the clinical examination, image diagnosis, and psychological state of patients with gastric cancer, and it is easy for patients to undergo image diagnosis and examination according to correct operating procedures.

Expression and role of nuclear receptor-interacting protein 1 (NRIP1) in stomach adenocarcinoma.

BackgroundNuclear receptor-interacting protein 1 (NRIP1), also named NR140, has been observed differentially express in multiple cancers, but the expression levels and the prognostic role of NRIP1 in stomach adenocarcinoma (STAD) remain unclear.MethodsWe used the Gene Expression Profiling Interactive Analysis (GEPIA) to analyze the NRIP1 expression levels in STAD, subgroups analysis of expression of NRIP1 via the UALCAN dataset. Further, cBioPortal was used to investigate the aberration type, co-mutations status, and located mutation of NRIP1. Correlated genes, and kinases, microRNA (miRNA), and transcription factor (TF) targets were identified using LinkedOmics. The Kaplan-Meier (K-M) plotter was used to analyze the prognosis of NRIP1 and the significantly correlated genes in STAD. Then, the tumor immune estimation resource (Timer) was used to explore the relation between NRIP1 and the immune cell infiltration, and the role of immune cells in STAD. The Human Protein Atlas (HPA) was used to confirm the NRIP1 protein express in STAD stomach tissue and normal stomach tissue.ResultsNRIP1 significantly overexpress in STAD, and the NRIP1 expression levels were impacted by clinical features. Overexpression of NRIP1 indicated the poor prognosis of STAD. Functional enrichment analysis showed the NRIP1 mainly enriched in immune response-regulating signaling pathway, cell-substrate adhesion, mRNA processing, and pathway in cancer. Overexpression USP25, SNYJ1 indicated the poor outcome of STAD, but the overexpression of BACH1 indicated protective biomarker. MIR-331 and MIR-132 have important role in STAD. Further, NRIP1 had a significant relation with immune infiltrates and other defined genes that significantly impact immune infiltrates. Immunohistochemical showed NRIP1 protein was higher in STAD than normal sample.ConclusionsIn this study, we revealed that overexpression of NRIP1 in the STAD sample compared to normal samples, NRIP1 significantly associated with macrophage. The high expression levels of NRIP1 and more macrophage infiltration led to poor prognosis of STAD.

S1992 Immune Checkpoint Inhibitor-Mediated Eosinophilic Esophagitis and Gastroenteritis

INTRODUCTION: Immune checkpoint inhibitors (ICPI) are immunomodulatory antibodies that enhance the immune system in the management of malignancies resulting in significantly improved prognosis especially in melanoma treatment. They are often associated with immune mediated side effects affecting multiple organ systems. The most common gastrointestinal side effect is colitis resulting in diarrhea, however eosinophilic esophagitis (EoE) and gastroenteritis is rarely reported. CASE DESCRIPTION/METHODS: A 50 year old female with history of stage IIIB melanoma managed with resection and 12 months of adjuvant nivolumab presented after immunotherapy completion with severe pruritis, dysphagia, and epigastric pain. Laboratory workup was notable for 19.9% eosinophilia with an absolute count of 1,700 cells/µL. Esophagogastroduodenoscopy (EGD) revealed ringed esophagus, white plaques and edema in the entire esophagus as well as edema and friability in the stomach and duodenal bulb. Biopsies showed increased intraepithelial eosinophils (eos), greater than 50 per high power field (hpf) in the proximal and distal esophagus. Gastric and duodenal biopsies revealed increased eos in the lamina propria with villous blunting. Her abdominal pain improved with a prednisone taper but she then developed severe heartburn and worsening dysphagia. Repeat EGD showed longitudinal furrows, edema, rings, and exudates consistent with EoE and normal stomach and duodenum. Pathology showed up to 80 eos/hpf in the esophagus, with resolution of the gastric and duodenal eosinophilia. Her symptoms resolved with a course of omeprazole and viscous budesonide slurry. DISCUSSION: Nivolumab is associated with several immune mediated toxicities including pneumonitis, colitis, endocrine and dermatological side effects. Eosinophilia may be seen following treatment with ICPIs and is potentially associated with improved prognosis. Eosinophilic tissue inflammation, particularly involving the GI tract is uncommon. A prior case report described eosinophilic enteritis following combination ICPIs ipilumab and nivolumab. A French case series reported 37 patients with hypereosinophilia following ICPI therapy; 57% had organ involvement, none with esophagitis or gastroenteritis. Most patients responded well to ICPI discontinuation and/or corticosteroids. This is the first report of eosinophilic esophagitis and gastroduodenitis following completion of 1 year of nivolumab therapy. There appears to be resolution with systemic and topical corticosteroid therapy.

Identification of miPEP133 as a novel tumor-suppressor microprotein encoded by miR-34a pri-miRNA

BackgroundVery few proteins encoded by the presumed non-coding RNA transcripts have been identified. Their cellular functions remain largely unknown. This study identifies the tumor-suppressor function of a novel microprotein encoded by the precursor of miR-34a. It consists of 133 amino acid residues, thereby named as miPEP133 (pri-microRNA encoded peptide 133).MethodsWe overexpressed miPEP133 in nasopharyngeal carcinoma (NPC), ovarian cancer and cervical cancer cell lines to determine its effects on cell growth, apoptosis, migration, or invasion. Its impact on tumor growth was evaluated in a xenograft NPC model. Its prognostic value was analyzed using NPC clinical samples. We also conducted western blot, immunoprecipitation, mass spectrometry, confocal microscopy and flow cytometry to determine the underlying mechanisms of miPEP133 function and regulation.ResultsmiPEP133 was expressed in normal human colon, stomach, ovary, uterus and pharynx. It was downregulated in cancer cell lines and tumors. miPEP133 overexpression induced apoptosis in cancer cells and inhibited their migration and invasion. miPEP133 inhibited tumor growth in vivo. Low miPEP133 expression was an unfavorable prognostic marker associated with advanced metastatic NPC. Wild-type p53 but not mutant p53 induced miPEP133 expression. miPEP133 enhanced p53 transcriptional activation and miR-34a expression. miPEP133 localized in the mitochondria to interact with mitochondrial heat shock protein 70kD (HSPA9) and prevent HSPA9 from interacting with its binding partners, leading to the decrease of mitochondrial membrane potential and mitochondrial mass.ConclusionmiPEP133 is a tumor suppressor localized in the mitochondria. It is a potential prognostic marker and therapeutic target for multiple types of cancers.

Abstract 5386: In situ detection of CLDN18.2 RNA in circulating tumor cells in gastrointestinal cancer

Abstract Objective: Recent clinical trials considered Claudin18 splice variants 2 (CLDN18.2) as the promising potential target for gastrointestinal(GI) cancer treatment. CLDN18.2 specifically express in normal stomach and gastric cancer, while ectopic activation can also be found among esophageal, pancreatic and bile duct cancers. However, high-consistency of CLDN18.1 and CLDN18.2 sequences makes it challenging to explore CLDN18.2-specific antibody. Therefore, detecting CLDN18.2 RNA expression may be another choice. Circulating tumor cells (CTCs) are regarded as biomarkers guiding treatment and tools investigated drug-resistant mechanisms. We aim to establish a CLDN18.2 RNA detection method in CTCs(CLDN18.2+ CTCs)for further application in monitoring anti-CLDN18.2 treatment of GI tumor. Methods: We established a CLDN18.2 RNA detection method based on designing a CLDN18.2-specific RNA molecular beacon (MB). 12 late stage GI cancer patients(pts)were enrolled. Paired blood (6ml) and tumor tissue specimens were respectively collected from 2 pancreatic and 10 gastric cancer pts. We evaluated CLDN18.2 expression on tissue by immunohistochemical (IHC) results. Then we analyzed CLDN18.2 expression on CTCs and tumor tissue. Results: Molecular and cell-line validation indicated that CLDN18.2 MB could specifically hybridized with CLDN18.2 RNA. In our study, Every pt had an evaluable CLDN18.2 IHC result. CTCs were identified in all of the pts (CTCs numbers ranging from 2 to 25). 9 of 12 pts had detectable CLDN18.2+CTCs, among which the CLDN18.2+CTCs ratio (CLDN18.2+CTCs/ Total number of CTCs) fluctuated from 14% to 100%. 96 CTCs fluorescence intensity were calculated statistically and classified by Agglomerative Clustering arithmetic. Consequently, we classified 3 clusters with significant differences (p<0.01). The cutoff points were 450rfu and 900rfu respectively. Hence, we subdivided CLDN18.2+CTCs into CLDN18.22+CTC and CLDN18.21+ CTC. Spearman Rank Correlation analysis showed that CLDN18.2+ CTCs ratio was concordant with IHC results. (P=0.007, Spearman=0.75). Conclusions: CLDN18.2 RNA positive CTCs ratio was consistent with CLDN18.2 expression level in GI cancer pts. It is warranted to further exploring the phenotypes and distinct mechanisms of CLDN18.2 RNA positive CTCs dynamic changes during anti-CLDN18.2 therapy. CLDN18.2 Expression in 12 GI cancer ptsPtsCancerCLDN18.2 Expression on Tissue(IHC)CLDN18.2 Expression in CTCs(CLDN18.2 MB)CLDN18.2-CLDN18.21+CLDN18.22+CTCs SumCLDN18.2+CTC ratio1GC020020%2GC0010010100%3GC040040%4GC030030%5GC2+07714100%6GC2+0257100%7PC2+410520%8PC2+0628100%9GC2+710814%10GC3+023225100%11GC3+221560%12GC3+0145100%*GC: Gastric cancer ; PC: Pancreatic cancer Citation Format: Xiaoyi Chong, Linyang Fan, Changsong Qi, Jifang Gong, Zhi Peng, Dan Liu, Yang Chen, Chenhui Xu, Yanyan Li, Congcong Ji, Qin Li, Zhiyuan Hu, Xiaotian Zhang, Lin Shen. In situ detection of CLDN18.2 RNA in circulating tumor cells in gastrointestinal cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5386.

Construction and Analysis of the Tumor-Specific mRNA-miRNA-lncRNA Network in Gastric Cancer.

Weighted correlation network analysis (WGCNA) is a statistical method that has been widely used in recent years to explore gene co-expression modules. Competing endogenous RNA (ceRNA) is commonly involved in the cancer gene expression regulation mechanism. Some ceRNA networks are recognized in gastric cancer; however, the prognosis-associated ceRNA network has not been fully identified using WGCNA. We performed WGCNA using datasets from The Cancer Genome Atlas (TCGA) and the Genotype-Tissue Expression (GTEx) to identify cancer-associated modules. The criteria of differentially expressed RNAs between normal stomach samples and gastric cancer samples were set at the false discovery rate (FDR) < 0.01 and |fold change (FC)| > 1.3. The ceRNA relationships obtained from the RNAinter database were examined by both the Pearson correlation test and hypergeometric test to confirm the mRNA–lncRNA regulation. Overlapped genes were recognized at the intersections of genes predicted by ceRNA relationships, differentially expressed genes, and genes in cancer-specific modules. These were then used for univariate and multivariate Cox analyses to construct a risk score model. The ceRNA network was constructed based on the genes in this model. WGCNA-uncovered genes in the green and turquoise modules are those most associated with gastric cancer. Eighty differentially expressed genes were observed to have potential prognostic value, which led to the identification of 12 prognosis-related mRNAs (KIF15, FEN1, ZFP69B, SP6, SPARC, TTF2, MSI2, KYNU, ACLY, KIF21B, SLC12A7, and ZNF823) to construct a risk score model. The risk genes were validated using the GSE62254 and GSE84433 datasets, with 0.82 as the universal cutoff value. 12 genes, 12 lncRNAs, and 35 miRNAs were used to build a ceRNA network with 86 dysregulated lncRNA–mRNA ceRNA pairs. Finally, we developed a 12-gene signature from both prognosis-related and tumor-specific genes, and then constructed a ceRNA network in gastric cancer. Our findings may provide novel insights into the treatment of gastric cancer.

Integration of GWAS and eQTL Analysis to Identify Risk Loci and Susceptibility Genes for Gastric Cancer.

Genome-wide association studies (GWAS) have identified several susceptibility loci for gastric cancer (GC), but the majority of identified single-nucleotide polymorphisms (SNPs) fall within the non-coding region and are likely to exert their biological function by modulating gene expression. To systematically estimate expression-associated SNPs (eSNPs) that confer genetic predisposition to GC, we evaluated the associations of 314,203 stomach tissue-specific eSNPs with GC risk in three GWAS datasets (2,631 cases and 4,373 controls). Subsequently, we conducted a gene-based analysis to calculate the cumulative effect of eSNPs through sequence kernel association combined test and Sherlock integrative analysis. At the SNP-level, we identified two novel variants (rs836545 at 7p22.1 and rs1892252 at 6p22.2) associated with GC risk. The risk allele carriers of rs836545-T and rs1892252-G exhibited higher expression levels of DAGLB (P = 3.70 × 10–18) and BTN3A2 (P = 3.20 × 10–5), respectively. Gene-based analyses identified DAGLB and FBXO43 as novel susceptibility genes for GC. DAGLB and FBXO43 were significantly overexpressed in GC tissues than in their adjacent tissues (P = 5.59 × 10–7 and P = 3.90 × 10–6, respectively), and high expression level of these two genes was associated with an unfavorable prognosis of GC patients (P = 1.30 × 10–7 and P = 7.60 × 10–3, respectively). Co-expression genes with these two novel genes in normal stomach tissues were significantly enriched in several cancer-related pathways, including P53, MAPK and TGF-beta pathways. In summary, our findings confirm the importance of eSNPs in dissecting the genetic basis of GC, and the identified eSNPs and relevant genes will provide new insight into the genetic and biological basis for the mechanism of GC development.

Gastric Microbiota in a Low-Helicobacter pylori Prevalence General Population and Their Associations With Gastric Lesions.

INTRODUCTION:Non–Helicobacter pylori microbiota might account for some cases with unexplained chronic gastritis that may in a minority eventually progress to gastric cancer through the Correa cascade. We characterized gastric microbiota by describing the normal stomach, compared it with early precancerous lesions and other disease states, and assessed whether H. pylori status affects bacterial diversity.METHODS:In a population-based study of those with and without gastrointestinal symptoms, cytology brush samples were collected during endoscopy from 316 individuals. Mucosal status was classified as normal mucosa (171), nonatrophic H. pylori gastritis (33), atrophic gastritis (12), or antral chemical gastritis (61). The 16S rRNA gene sequencing and analysis were performed to characterize the microbiota.RESULTS:Microbiota in atrophic gastritis and nonatrophic H. pylori gastritis stomachs were dysbiotic and differed from those in the normal stomach (P = 0.001). The normal stomach had the highest microbial diversity, followed by antral chemical gastritis. The atrophic gastritis and chronic H. pylori gastritis groups had the lowest diversity, a difference that was statistically significant (P = 0.01). Besides H. pylori, non–H. pylori bacteria accounted for group differences. Microbial network analysis showed that the normal group network was most highly connected, whereas the H. pylori gastritis group had the lowest connection. We found an increasing positive co-occurrence of oral bacteria in the stomach because samples deviated from the normal network, some of which were pathogens. The H. pylori–negative group had the highest microbial diversity (Shannon index) compared with the H. pylori–positive group (P = 0.001).DISCUSSION:In this low–H. pylori prevalence general population, the gastric mucosal microbiota of the normal stomach differed significantly from those with nonatrophic or atrophic gastritis. There was an increasing abundance of pathogenic bacteria from the normal state to early precancerous states.

Identification of critical genes in gastric cancer to predict prognosis using bioinformatics analysis methods.

Ranking fourth in the world in tumor incidence and second in cancer-related death worldwide, gastric cancer (GC) is one of the major malignant tumors, and has a very complicated pathogenesis. In the present study, we aimed to identify new biomarkers to predict the survival rate of GC patients. The differentially expressed genes (DEGs) between GC tissues and normal stomach tissues were obtained by using GEO2R, and overlapped DEGs were acquired with Venn diagrams. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis were conducted with R software. Then, the protein-protein interaction (PPI) of these DEGs was visualized by Cytoscape. Gene Expression Profiling Interactive Analysis (GEPIA) was used to verify the expression differences of hub genes in gastric adenocarcinoma tissues and normal tissues. Overall survival (OS) of hub genes was calculated by Kaplan-Meier plotter. There were a total of 128 consistently expressed genes in the two datasets: 85 upregulated genes were enriched in extra-cellular matrix (ECM)-receptor interaction, protein digestion and absorption, focal adhesion, gastric acid secretion, mineral absorption, systemic lupus erythematosus, amoebiasis, and PI3K-Akt signaling pathway, and 43 downregulated genes were enriched in palate development, blood coagulation, positive regulation of transcription from RNA polymerase II promoter, axonogenesis, receptor internalization, negative regulation of transcription from RNA polymerase II promoter, and in no significant signaling pathways. From the PPI network analyzed by Molecular Complex Detection (MCODE) plug-in, all 27 upregulated genes were selected. Furthermore, to analyze the OS among these genes, Kaplan-Meier analysis was conducted, and 25 genes were associated with remarkably worse survival. For validation in GEPIA, 11 of 25 genes were discovered to be highly expressed in GC tissues compared to normal OS tissues. Furthermore, in the re-analysis of the Database for Annotation, Visualization and Integrated Discovery (DAVID), three genes [G2/miotic-specific cyclin B1 (CCNB1), polo-like kinases 1 (PLK1), and pituitary tumor-transforming gene-1 (PTTG1)] were markedly enriched in the cell cycle pathway, particulary the G1-G1/S phase. Three remarkably upregulated DEGs with poor prognosis in GC were identified and may serve as new prognostic biomarkers and targets in GC therapy.

FB-15 inhibits MGC-803 cells growth by regulating energy metabolism

In this study, we scrutinized the anticancer effects of FB-15 on human gastric carcinoma MGC-803 cells in vitro and vivo, and its preliminary effect on tubulin and HIF-1α. We confirmed that FB-15 not only inhibited the proliferation of a large number of cells in a concentration and time-dependent manner but also inhibited proliferation of a single cell to form clones. FB-15 manifested little cytotoxicity for normal stomach cells GES-1. The flow cytometry analysis displayed that FB-15 induced apoptosis MGC-803 cells and mainly arrested cells in the S phase in a concentration-dependent manner. The results of the wound healing assay indicated that FB-15 suppressed cell migration. Furthermore, the western blotting showed that FB-15 down-regulated the expression of β3-tubulin and HIF-1α, consistent with Immunohistochemical assay. The binding modes of FB-15 with tubulin were clarified by molecular docking. FB-15 significantly suppressed the growth of MGC-803 gastric cancer tumors. The inhibitory effect of FB-15 on tumor growth was superior to 5-Fu. Taken together, these results provided evidence for FB-15 to be used as an effective anticancer drug candidate for gastric cancer.

Incisura angularis belongs to fundic or transitional gland regions in Helicobacter pylori-naïve normal stomach: Sub-analysis of the prospective multi-center study.

Incisura angularis is one of the important parts for evaluating mucosal atrophy and cancer risk. We determined the type of mucosa at incisura angularis in Helicobacter pylori-naïve normal stomach. Subjects aged 40years or older who underwent esophagogastroduodenoscopy for dyspepsia or a routine health checkup were recruited in 24 facilities between March 2008 and February 2009. Serum antibody to H. pylori was measured. Endoscopic atrophy was evaluated according to Updated Kimura-Takemoto classification. Five biopsy specimens were taken from the incisura angularis and greater and lesser curvatures of the antrum and corpus. These specimens were histologically classified as fundic, pyloric or transitional. H.pylori-naïve normal stomach was defined with the strictest criterion among various combinations of histological, endoscopic and serum findings. We determined histological type of mucosa at incisura angularis in H.pylori-naïve normal stomach. A total of 270 subjects (122 men, mean 64.6yo) were analyzed. The strictest criterion consists of serum antibody≤3.0U/mL, endoscopic atrophy C-1 and histological grade 0 in all of the five items in Updated Sydney System. The numbers having fundic, transitional and pyloric mucosa at incisura angularis under the strictest criterion were 13 (50%), 13 (50%) and 0, respectively. The probability that the type of mucosa at incisura angularis would be pyloric was almost zero (97.5% confidence interval 0-0.132). Incisura angularis of the stomach may not belong to pyloric, but fundic or transitional mucosa in H.pylori-naïve normal stomach. UMIN000018218.

Downregulation of PHF19 inhibits cell growth and migration in gastric cancer

Objectives: The PHD Finger Protein 19 (PHF19), as a sub-component of polycomb repressive complex 2 (PRC2), has been identified to be associated with various biological processes. Aberrant expression of PHF19 has implicated in several cancer types. This study aims to investigate its function and clinical significance in gastric cancer for the first time.Methods: The expression of PHF19 was evaluated by quantitative real-time PCR (qRT-PCR) and immunohistochemistry. PHF19 was silenced by small interference RNAs and lentiviral particles in gastric cancer cells. Then cell growth was measured by CCK-8 assays, colony formation and in a mouse model. Transwell and wound healing assays were performed to detect cell migration. Western blot analysis was used to explore the downstream signaling factors in PHF19-silenced cells, xenograft tumors and gastric cancer samples.Results: PHF19 was frequently upregulated in gastric cancer tissues compared with adjacent normal stomach tissues and this upregulation was correlated with tumor cell differentiation and poor outcome of gastric cancer patients. Functionally, the silencing of PHF19 in gastric cancer cells led to decreased cell growth and migration. Stable knockdown of PHF19 inhibited the tumorigenicity of gastric cancer cells in nude mice model. Western blot results demonstrated that phosphorylated AKT and ERK were reduced upon PHF19 downregulation, implying the two signaling pathways possibly mediate the oncogenic roles of PHF19.Conclusions: We identified PHF19 as an oncogene candidate and provided a new potential drug target for gastric cancer.

NFKB2 gene expression in patients with peptic ulcer diseases and gastric cancer

Gastric cancer is one of the most common worldwide types of cancer. It is a multifactorial disease and both environmental and genetic factors play an important role in its etiology. Evaluation of the relative expression level of NFKB2 gene in two groups of patients: peptic ulcer and gastric cancer and its role in the pathomechanism of these diseases was the aim of this study. RNA was isolated from: 79 samples of peptic ulcer, 22 gastric cancer and 11 control tissue. The real-time PCR technique was used to study the expression of NFKB2 gene. The relative expression level of NFKB2 gene was a variable in all three studied groups. The relative NFKB2 gene expression depends on the type of a disease. Peptic ulcer cases showed the increased relative NFKB2 gene expression to control group (p = 0.0000). Cancer cases presented decreased relative NFKB2 gene expression to normal stomach tissue (p = 0.0183). There are statistically important differences in the investigated gene expression between peptic ulcer, where the expression level is higher comparing to gastric cancer and control tissue which confirmed that such an activation is connected with an inflammatory process. The relative expression level of NFKB2 is decreased in cancer cases as opposed to control tissue and peptic ulcer cases which could suggest that during carcinogenesis of gastric cancer inhibition of NF-kB pathway takes place which could be a promising factor for patients.