Effects of long-term aspirin use on molecular alterations in precancerous gastric mucosa in patients with and without gastric cancer.
Michigami Yuki,Watari Jiro,Ito Chiyomi,Hara Ken,Yamasaki Takahisa,Kondo Takashi,Kono Tomoaki,Tozawa Katsuyuki,Tomita Toshihiko,Oshima Tadayuki,Fukui Hirokazu,Morimoto Takeshi,Das Kiron M,Miwa Hiroto
The risk of gastric cancer (GC) remains even after H. pylori eradication; thus, other combination treatments, such as chemopreventive drugs, are needed. We evaluated the effects of aspirin on genetic/epigenetic alterations in precancerous conditions, i.e., atrophic mucosa (AM) and intestinal metaplasia (IM), in patients with chronic gastritis who had taken aspirin for more than 3 years. A total of 221 biopsy specimens from 74 patients, including atrophic gastritis (AG) cases without aspirin use (control), AG cases with aspirin use (AG group), and GC cases with aspirin use (GC group), were analyzed. Aspirin use was associated with a significant reduction of CDH1 methylation in AM (OR: 0.15, 95% CI: 0.06-0.41, p = 0.0002), but was less effective in reversing the methylation that occurred in IM. Frequent hypermethylation including that of CDH1 in AM increased in the GC group compared to the AG group, and CDH1 methylation was an independent predictive marker of GC (OR: 8.50, 95% CI: 2.64-25.33, p = 0.0003). In patients with long-term aspirin use, the changes of molecular events in AM but not IM may be an important factor in the reduction of cancer incidence. In addition, methylation of the CDH1 gene in AM may be a surrogate of GC.
Analysis of correlation between HP infection and activation of PI3K/Akt pathway in mucosal tissues of gastric cancer and precancerous lesions.
Xie Yi,Liu Li
The aim of the study was to investigate the correlation between (HP) infection and activation of the phosphatidylinositol-3-kinase/Akt (PI3K/Akt) pathway in mucosal tissues of gastric cancer and precancerous lesions. Patients with chronic atrophic gastritis (n=52) and gastric cancer (n=98) were treated in the Department of Gastroenterology at The Fifth People's Hospital of Chongqing from August 2011 to August 2016 were selected, and the biopsy tissue and serum specimens were collected. The HP infection was detected via enzyme-linked immunosorbent assay (ELISA), and the expression level of phosphorylated-Akt (p-Akt) was detected via immunohistochemistry (IHC). Moreover, experiments were performed to simulate HP infection in gastric cancer cells (MGC-803 and AGS), and the p-Akt protein level, PI3K activity and cell proliferative activity were detected. Finally, the changes in Akt protein level were detected by co-culture of gastric cancer cells via LY294002, a PI3K inhibitor, and HP. The positive rate of HP infection in patients with chronic atrophic gastritis was 84.6% (44/52), which was significantly higher than that in patients with gastric cancer [73.5% (72/98)] (p<0.05). The positive rate of HP infection in patients with early gastric cancer (86.4%) was significantly higher than that in patients with moderate-advanced gastric cancer (69.7%) (p<0.05). Results of IHC and western blot analysis revealed that the p-Akt expression level in HP-positive tissues was obviously higher than that in HP-negative tissues (p<0.05). cell experiments revealed that the PI3K activity was enhanced and the PI3K/Akt pathway was significantly activated after HP infection in tumor cells, thus promoting the proliferation of tumor cells (p<0.05) in a time-dependent manner. After LY294002 inhibited PI3K activity, Akt was not significantly activated by HP infection. Thus, HP activates the PI3K/Akt pathway in gastric cancer cells, thereby promoting tumor cell proliferation.
Indoleamine 2,3-Dioxygenase 1, Increased in Human Gastric Pre-Neoplasia, Promotes Inflammation and Metaplasia in Mice and Is Associated With Type II Hypersensitivity/Autoimmunity.
El-Zaatari Mohamad,Bass Adam J,Bowlby Reanne,Zhang Min,Syu Li-Jyun,Yang Yitian,Grasberger Helmut,Shreiner Andrew,Tan Bei,Bishu Shrinivas,Leung Wai K,Todisco Andrea,Kamada Nobuhiko,Cascalho Marilia,Dlugosz Andrzej A,Kao John Y
BACKGROUND & AIMS:Chronic gastrointestinal inflammation increases the risk of cancer by mechanisms that are not well understood. Indoleamine-2,3-dioxygenase 1 (IDO1) is a heme-binding enzyme that regulates the immune response via catabolization and regulation of tryptophan availability for immune cell uptake. IDO1 expression is increased during the transition from chronic inflammation to gastric metaplasia. We investigated whether IDO1 contributes to the inflammatory response that mediates loss of parietal cells leading to metaplasia. METHODS:Chronic gastric inflammation was induced in Ido1 and CB57BL/6 (control) mice by gavage with Helicobacter felis or overexpression of interferon gamma in gastric parietal cells. We also performed studies in Jh mice, which are devoid of B cells. Gastric tissues were collected and analyzed by flow cytometry, immunostaining, and real-time quantitative polymerase chain reaction. Plasma samples were analyzed by enzyme-linked immunosorbent assay. Gastric tissues were obtained from 20 patients with gastric metaplasia and 20 patients without gastric metaplasia (controls) and analyzed by real-time quantitative polymerase chain reaction; gastric tissue arrays were analyzed by immunohistochemistry. We collected genetic information on gastric cancers from The Cancer Genome Atlas database. RESULTS:H felis gavage induced significantly lower levels of pseudopyloric metaplasia in Ido1 mice, which had lower frequencies of gastric B cells, than in control mice. Blood plasma from H felis-infected control mice had increased levels of autoantibodies against parietal cells, compared to uninfected control mice, but this increase was lower in Ido1 mice. Chronically inflamed stomachs of Ido1 mice had significantly lower frequencies of natural killer cells in contact with parietal cells, compared with stomachs of control mice. Jh mice had lower levels of pseudopyloric metaplasia than control mice in response to H felis infection. Human gastric pre-neoplasia and carcinoma specimens had increased levels of IDO1 messenger RNA compared with control gastric tissues, and IDO1 protein colocalized with B cells. Co-clustering of IDO1 messenger RNA with B-cell markers was corroborated by The Cancer Genome Atlas database. CONCLUSIONS:IDO1 mediates gastric metaplasia by regulating the B-cell compartment. This process appears to be associated with type II hypersensitivity/autoimmunity. The role of autoimmunity in the progression of pseudopyloric metaplasia warrants further investigation.
Methylation and Expression of Nonclustered Protocadherins Encoding Genes and Risk of Precancerous Gastric Lesions in a High-Risk Population.
Wu Si,Zhang Yang,Zhang Lian,Ma Jun-Ling,Zhou Tong,Li Zhe-Xuan,Liu Wei-Dong,Li Wen-Qing,You Wei-Cheng,Pan Kai-Feng
Cancer prevention research (Philadelphia, Pa.)
Nonclustered protocadherins (PCDH) family is a group of cell-cell adhesion molecules. We have found differentially methylated genes in the nonclustered PCDHs family associated with () infection in prior genome-wide methylation analysis. To further investigate the methylation and expression of nonclustered PCDHs encoding genes in -related gastric carcinogenesis process, four candidate genes including and were selected, which were reported to be tumor suppressors for digestive cancers. A total of 747 participants with a spectrum of gastric lesions were enrolled from a high-risk population of gastric cancer. Promoter methylation levels of four genes were significantly higher in positive subjects than the negative group (all < 0.001). Elevated methylation levels of and were observed with the increasing severity of gastric lesions (both < 0.001). In the protein expression analysis, PCDH17 expression was inversely associated with gastric lesions; the OR [95% confidence interval (CI)] was 0.49 (0.26-0.95) for chronic atrophic gastritis (CAG), 0.31 (0.15-0.63) for intestinal metaplasia, and 0.38 (0.19-0.75) for indefinite dysplasia and dysplasia, compared with superficial gastritis. In addition, PCDH10 expression was significantly lower in CAG (OR, 0.40; 95% CI, 0.24-0.68). The inverse association between methylation and protein expression of and 7 was further supported when we explored the methylation and mRNA expression in The Cancer Genome Atlas database (all < 0.001). Our study found elevated promoter methylation and decreased expression of and in advanced gastric lesions, suggesting that elevated and methylation may be an early event in gastric carcinogenesis. .
Associations among Gastric Juice pH, Atrophic Gastritis, Intestinal Metaplasia and Infection.
Sung Jihee,Kim Nayoung,Lee Jongchan,Hwang Young-Jae,Kim Hyoung Woo,Chung Jung Wha,Kim Jin-Wook,Lee Dong Ho
Gut and liver
Background/Aims:Gastric juice plays a crucial role in the physiology of the stomach. The aim of this study is to evaluate associations among the pH of gastric juice, atrophic gastritis (AG), intestinal metaplasia (IM), pepsinogen, and infection. Methods:Gastric biopsies and juice were collected from 46 subjects who underwent endoscopies at Seoul National University Bundang Hospital between November 2011 and March 2013. , AG and IM were evaluated, and pepsinogen I or II, I/II ratio, and interleukin (IL)-1β levels were measured. Results:The mean pH of gastric juice was higher in the -positive group (n=17) than that in the -negative group (n=29) (4.54 vs 2.46, p=0.002). When patients were divided into pH ＜3 (n=28) and pH ≥3 (n=18) groups, was lower in the pH ＜3 group (21.4%) than in the pH ≥3 group (61.1%) (p=0.007). The pH ≥3 group demonstrated AG and IM more frequently than the pH ＜3 group in the body (p=0.047 and p=0.051, respectively) but not in the antrum. There were no differences in pepsinogen I or II, I/II ratio, and IL-1β levels between the two groups. Conclusions:There is a relationship between chronic infection and gastric juice pH ≥3, which may originate from AG and IM in the body.
Hepatoma-derived growth factor participates in Helicobacter Pylori-induced neutrophils recruitment, gastritis and gastric carcinogenesis.
Chu Tian-Huei,Huang Shih-Tsung,Yang Sheau-Fang,Li Chia-Jung,Lin Hung-Wei,Weng Bi-Chuang,Yang Shih-Ming,Huang Shih-Chung,Wu Jian-Ching,Chang Yi-Chen,Wen Zhi-Hong,Chen Yi-Ming Arthur,Wu Wen-Jeng,Kung Mei-Lang,Tai Po-Han,Wu Deng-Chyang,Tai Ming-Hong
Helicobacter pylori (Hp) infection and overexpression of hepatoma-derived growth factor (HDGF) are involved in gastric carcinogenesis. However, the relationship between Hp-induced gastric diseases and HDGF upregulation is not yet completely clear. This study aimed to elucidate the role of HDGF in Hp-induced gastric inflammation and carcinogenesis. HDGF expression in gastric biopsy and serum from patients was analyzed by immunohistochemical and ELISA analysis, respectively. Hp and gastric cells coculture system was employed to delineate the mechanism underlying HDGF overexpression during Hp infection. The gastric pathologies of wild type and HDGF knockout mice after Hp infection were investigated by immunohistochemical, immunoblot, and immunofluorescence analyses. HDGF level was significantly elevated in patients with Hp infection or intestinal metaplasia (IM, a precancerous lesion), and HDGF overexpression was positively correlated with Hp load, IM, and neutrophil infiltration in gastric biopsy. Consistently, patients with Hp infection or IM had significantly higher serum HDGF level. By using coculture assay, Hp infection led to HDGF upregulation and secretion in gastric cells. In mice model, HDGF ablation significantly suppressed the Hp-induced neutrophil infiltration and inflammatory TNF-α/COX-2 signaling, thereby relieving the tissue damage in stomach. This was further supported by that recombinant HDGF (rHDGF) stimulated the differentiation/chemotaxis of cultured neutrophils and oncogenic behaviors of gastric cells. Time series studies showed that Hp infection elicited an inflammatory TNF-α/HDGF/COX-2 cascade in stomach. HDGF secretion by Hp infection promotes the neutrophils infiltration and relays Hp-induced inflammatory signaling. Thus, HDGF may constitute a novel diagnostic marker and therapeutic target for Hp-induced gastritis and carcinogenesis.
Oxidative Phosphorylation System in Gastric Carcinomas and Gastritis.
Feichtinger René G,Neureiter Daniel,Skaria Tom,Wessler Silja,Cover Timothy L,Mayr Johannes A,Zimmermann Franz A,Posselt Gernot,Sperl Wolfgang,Kofler Barbara
Oxidative medicine and cellular longevity
Switching of cellular energy production from oxidative phosphorylation (OXPHOS) by mitochondria to aerobic glycolysis occurs in many types of tumors. However, the significance of this switching for the development of gastric carcinoma and what connection it may have to infection of the gut, a primary cause of gastric cancer, are poorly understood. Therefore, we investigated the expression of OXPHOS complexes in two types of human gastric carcinomas ("intestinal" and "diffuse"), bacterial gastritis with and without metaplasia, and chemically induced gastritis by using immunohistochemistry. Furthermore, we analyzed the effect of HP infection on several key mitochondrial proteins. Complex I expression was significantly reduced in intestinal type (but not diffuse) gastric carcinomas compared to adjacent control tissue, and the reduction was independent of HP infection. Significantly, higher complex I and complex II expression was present in large tumors. Furthermore, higher complex II and complex III protein levels were also obvious in grade 3 versus grade 2. No differences of OXPHOS complexes and markers of mitochondrial biogenesis were found between bacterially caused and chemically induced gastritis. Thus, intestinal gastric carcinomas, but not precancerous stages, are frequently characterized by loss of complex I, and this pathophysiology occurs independently of HP infection.
Tongue coating microbiome as a potential biomarker for gastritis including precancerous cascade.
Cui Jiaxing,Cui Hongfei,Yang Mingran,Du Shiyu,Li Junfeng,Li Yingxue,Liu Liyang,Zhang Xuegong,Li Shao
Protein & cell
The development of gastritis is associated with an increased risk of gastric cancer. Current invasive gastritis diagnostic methods are not suitable for monitoring progress. In this work based on 78 gastritis patients and 50 healthy individuals, we observed that the variation of tongue-coating microbiota was associated with the occurrence and development of gastritis. Twenty-one microbial species were identified for differentiating tongue-coating microbiomes of gastritis and healthy individuals. Pathways such as microbial metabolism in diverse environments, biosynthesis of antibiotics and bacterial chemotaxis were up-regulated in gastritis patients. The abundance of Campylobacter concisus was found associated with the gastric precancerous cascade. Furthermore, Campylobacter concisus could be detected in tongue coating and gastric fluid in a validation cohort containing 38 gastritis patients. These observations provided biological evidence of tongue diagnosis in traditional Chinese medicine, and indicated that tongue-coating microbiome could be a potential non-invasive biomarker, which might be suitable for long-term monitoring of gastritis.
Prostaglandin E induces DNA hypermethylation in gastric cancer and .
Wong Chi Chun,Kang Wei,Xu Jiaying,Qian Yun,Luk Simson Tsz Yat,Chen Huarong,Li Weilin,Zhao Liuyang,Zhang Xiaoming,Chiu Phlip Wy,Ng Enders Kw,Yu Jun
: Prostaglandin E (PGE) is a pro-inflammatory eicosanoid up-regulated in gastric cancer (GC). However, its impact on epigenetic dysfunction in the process of gastric carcinogenesis is unknown. In this study, we investigate the role of PGE in DNA methylation in gastric epithelium , in mice, and humans. : PGE-induced DNMT3B and DNA methylation was determined in gastric cell lines and COX-2 transgenic mice. Effect of COX-2 inhibition on DNA methylation was evaluated in a randomized controlled trial. Efficacy of combined COX-2/PGE and DNMT inhibition on GC growth was examined in cell lines and mice models. : PCR array analysis of PGE-treated GC cells revealed the up-regulation of DNMT3B, a DNA methyltransferase. In GC cells, PGE induced DNMT3B expression and activity, leading to increased methylated cytosine (5mC) and promoter methylation of tumor suppressive genes (MGMT and CNR1). Consistently, Cox-2 (rate-limiting enzyme for PGE biosynthesis) transgenic expression in mice significantly induced Dnmt3b expression, increased 5mC content, and promoted Mgmt promoter methylation. We retrospectively analyzed the 5mC content of 42 patients with intestinal metaplasia (a precancerous lesion of GC) treated with a COX-2 specific inhibitor Rofecoxib or placebo for 2 years, revealing that the COX-2 inhibitor significantly down-regulated 5mC levels (N=42, P=0.009). Collectively, these data indicate that PGE is closely related to DNA hypermethylation and . Using genome-wide 450K methylation array, we identified chromosomal genes (POT1, ATM and HIST1H2AA) were preferentially methylated by PGE. Biofunctional work revealed that POT1 functions as a tumor suppressor. Finally, we demonstrated that combinatorial inhibition of COX-2 and DNMT using Celecoxib and Decitabine synergistically inhibited GC growth and . : This study suggested that PGE promotes DNA methylation in GC, and that co-targeting of PGE and DNMT inhibits GC.
NMR-based metabonomics and correlation analysis reveal potential biomarkers associated with chronic atrophic gastritis.
Cui Jiajia,Liu Yuetao,Hu Yinghuan,Tong Jiayu,Li Aiping,Qu Tingli,Qin Xuemei,Du Guanhua
Journal of pharmaceutical and biomedical analysis
Chronic atrophic gastritis (CAG) is one of the most important pre-cancerous states with a high prevalence. Exploring of the underlying mechanism and potential biomarkers is of significant importance for CAG. In the present work, H NMR-based metabonomics with correlative analysis was performed to analyze the metabolic features of CAG. 19 plasma metabolites and 18 urine metabolites were enrolled to construct the circulatory and excretory metabolome of CAG, which was in response to alterations of energy metabolism, inflammation, immune dysfunction, as well as oxidative stress. 7 plasma biomarkers and 7 urine biomarkers were screened to elucidate the pathogenesis of CAG based on the further correlation analysis with biochemical indexes. Finally, 3 plasma biomarkers (arginine, succinate and 3-hydroxybutyrate) and 2 urine biomarkers (α-ketoglutarate and valine) highlighted the potential to indicate risks of CAG in virtue of correlation with pepsin activity and ROC analysis. Here, our results paved a way for elucidating the underlying mechanisms in the development of CAG, and provided new avenues for the diagnosis of CAG and presented potential drug targets for treatment of CAG.
Short-form RON (sf-RON) enhances glucose metabolism to promote cell proliferation via activating β-catenin/SIX1 signaling pathway in gastric cancer.
Wang Ziliang,Yang Yufei,Hu Shuang,He Jian,Wu Zheng,Qi Zihao,Huang Mingzhu,Liu Rujiao,Lin Ying,Tan Cong,Xu Midie,Zhang Zhe
Cell biology and toxicology
Recepteur d'origine nantais (RON) has been implicated in cell proliferation, metastasis, and chemoresistance of various human malignancies. The short-form RON (sf-RON) encoded by RON transcripts was overexpressed in gastric cancer tissues, but its regulatory functions remain illustrated. Here, we found that sf-RON promoted gastric cancer cell proliferation by enhancing glucose metabolism. Furthermore, sf-RON was proved to induce the β-catenin expression level through the AKT1/GSK3β signaling pathway. Meanwhile, the binding sites of β-catenin were identified in the promoter region of SIX1 and it was also demonstrated that β-catenin positively regulated SIX1 expression. SIX1 enhanced the promoter activity of key proteins in glucose metabolism, such as GLUT1 and LDHA. Results indicated that sf-RON regulated the cell proliferation and glucose metabolism of gastric cancer by participating in a sf-RON/β-catenin/SIX1 signaling axis and had significant implications for choosing the therapeutic target of gastric cancer.
CagA Protein Attenuates 5-Fu Sensitivity of Gastric Cancer Cells Through Upregulating Cellular Glucose Metabolism.
Gao Sujie,Song Defeng,Liu Yiting,Yan Hongwei,Chen Xuebo
OncoTargets and therapy
Introduction:Gastric cancer (GC) is one of the most malignancies leading to human mortality due to its development, progress, metastasis and poor prognosis, despite the development of remarkable chemotherapy and surgery. The 5-ﬂuorouracil (5-Fu) is an effective anti-gastric cancer agent. However, a fraction of GC patients acquire 5-Fu chemoresistance. Methods:In this study, the CagA protein was detected from CagA-positive gastric cancer patients by qRT-PCR and immunohistochemistry. The 5-Fu resistant gastric cancer cell line was generated from MKN45-CagA cells which was transfected with CagA overexpression vector. Cellular glucose metabolism was determined by measurements of glucose uptake, lactate product and glycolysis enzymes. Results:We report that the ()-secreted (CagA) oncoprotein is positively correlated with 5-Fu resistance of gastric cancer. From totally 72 CagA-positive gastric cancer patients, CagA high-expressed patients showed more resistance to 5-Fu than CagA low-expressed patients. Moreover, statistical analysis revealed that CagA mRNA and protein expressions were significantly upregulated in 5-Fu resistant gastric cancer patients. We observed that CagA protein is upregulated in 5-Fu resistant gastric cancer cells compared with sensitive cells. Interestingly, cellular glucose metabolism was upregulated; the glucose uptake and lactate production were significantly higher in 5-Fu resistant gastric cancer cells. The Akt phosphorylation and expressions of glycolysis key enzymes, Hexokinase 2 and LDHA, were significantly upregulated in 5-Fu resistant gastric cancer cells. On the other way, inhibition of glycolysis or Akt pathway effectively overcame 5-Fu resistance from both in vitro and in vivo models. Finally, we report that the combination of Akt or glycolysis inhibitor with 5-Fu could synergistically enhance the cytotoxicity of 5-Fu to CagA-overexpressed gastric cancer cells. Discussion:In summary, our study demonstrated a CagA-Akt-glycolysis-5-Fu resistance axis, contributing to the development of new therapeutic agents against chemoresistant human gastric cancer.