科创中国

Hypoxia is a common environmental stress that influences signaling pathways and cell function, which through initiates intracellular signaling pathways and hence leads to the activation of the transcription factor hypoxia-inducible factor 1 (HIF-1). In this study, we initially confirm that hypoxia activates HIF-1α protein expression in a time-dependent manner with a maximum reached at 60 min in vitro and 4h in vivo in gastric mucosa epithelial cells. The expression of HIF-1α is correlated with the activation of HIF-1 DNA binding and transcriptional activity. Hypoxia does not affect HIF-1α mRNA transcription but regulates HIF-1α protein expression through a translation-dependent pathway to regulate protein synthesis. Hypoxia could induce phosphorylation of Akt, MAPK (ERK), and a target of p70S6K1. PI3K and MAPK inhibitor and LY294002 and U0126 could inhibit hypoxia-induced HIF-1 and VEGF expression. We also investigated the role of reactive oxygen species (ROS) involved in HIF-1 and VEGF expression. Exogenous addition of H2O 2 was sufficient to activate Akt and ERK, scavengers of H 2O2 significantly inhibited hypoxia-induced Akt and ERK, and subsequent HIF-1α expression and transcriptional activity. In conclusion, our data suggested that hypoxia-PI3K signaling through Akt and ERK kinases regulated ROS-dependent, hypoxia-induced HIF-1 activation and VEGF expression in gastric mucosa epithelial cells. © 2008 MAIK Nauka.

Liu Lili;Ning Xiaoxuan;Han Shuang;Zhang Hongbo;Sun Li;Shi Yongquan;Sun Shiren;Guo Changcun;Yin Fang;Qiao Taidong;Wu Kaichun;Fan Daiming

Molecular Biology

2008

Background: Liver cirrhosis represents the end stage of chronic liver injury. Currently, liver transplantation provides the only definite cure but it is beset with many problems, including lack of donors and risk of rejection. Stem cell therapy is very attractive in this setting because it has the potential to help tissue regeneration. In this study, we aimed to investigate the therapeutic effect of peripheral blood monocyte cell (PBMC) transplantation in decompensated liver cirrhosis. Methods: A total of 40 subjects (31 men and nine females, age range 21-71 years) was recruited to two groups. Group 1 received granulocyte-colony-stimulating factor (G-CSF) mobilization, PBMC collection by leukapheresis and PBMC transplant therapy. Group 2 received G-CSF mobilization for 4 days. At baseline and 6 months after treatment, liver function of the two groups was monitored by blood examination and ultrasonagraphy. Results: Both groups gained significant improvement in liver synthetic function, such as serum albumin and prothrombin time, from baseline to 6 months after treatment (P < 0.01). However, there was no significant difference in alanine aminotransferase, aspartate aminotransferase and total bilirubin in both groups (P > 0.05). Compared with group 2, a significantly improved liver function was observed in group 1, including elevated serum albumin level and a decreased CTP score (P < 0.05). No major adverse effects were noted. Discussion: Autologous PBMC transplantation could be considered as a novel and alternative treatment for patients with decompensated liver cirrhosis.

Han Y.;Yan L.;Han G.;Zhou X.;Hong L.;Yin Z.;Zhang X.;Wang J.;Sun A.;Liu Z.;Xie H.;Wu K.;Ding J.;Fan D.;Wang S.

Cytotherapy

2008

Calcyclin-binding protein (CacyBP)/Siah-1 interacting protein (SIP), a component of ubiquitin-mediated proteolysis, could bind the Skp1-Cul1-F box protein complex. Although CacyBP/SIP was implicated in p53-induced β-catenin degradation, its exact function was still unknown. Our previous studies showed that CacyBP/SIP could modulate the multidrug-resistant phenotype of gastric cancer cells and was highly expressed in gastric cancer tissues compared with that in non-cancerous tissues. In this study, CacyBP/SIP protein expression profile in a broad range of human normal tissues and carcinomas was analyzed by immunohistochemistry staining with anti-CacyBP/SIP monoclonal antibody first produced in our laboratory. CacyBP/SIP was generally localized in the cytoplasm/nucleus. Positive staining of CacyBP/SIP was found in brain, heart, lymph node, and esophagus. Weak staining was shown in the rectum and kidney. No CacyBP/SIP was detected in other normal tissues. However, CacyBP/SIP was ubiquitously detected in all kinds of tumor tissues and was highly expressed in nasopharyngeal carcinoma, osteogenic sarcoma, and pancreatic cancer. To our knowledge, this is the first study on the CacyBP/SIP expression pattern in a broad range of human normal and tumor tissues. The data presented should serve as a useful reference for other investigators in future studies of CacyBP/SIP functions. Hopefully, this knowledge will lead to discovery of more roles of CacyBP/SIP in tumorigenesis. © The Histochemical Society, Inc.

Zhai Huihong;Shi Yongquan;Jin Haifeng;Lu Yuanyuan;Chen Xiong;Wang Jinbo;Wang Xin;Fan Daiming M.;Li Yuanfei;Ding Liping

Journal of Histochemistry and Cytochemistry

2008

Hong Liu;Wu Kaichun;Fan Daiming

Annals of Internal Medicine

2008

Hypoxia is a common environmental stress that influences signaling pathways and cells function, which through initiating intracellular signaling pathways and hence leading to the activation of the transcription factor hypoxia-inducible factor-1 (HIF-1). In this study, we initially confirm that hypoxia activates HIF-1alpha protein expression in a time-dependent manner with a maximum reached at 60 min in vitro and 4h in vivo in gastric mucosa epithelial cells. The expression of HIF-1alpha is correlated with the activation of HIF-1 DNA binding and transcriptional activity. Hypoxia dose not affect HIF-1alpha mRNA transcription but regulates HIF-1alpha protein expression through a translation-dependent pathway to regulate protein synthesis. Hypoxia could induce phosphorylation of Akt, MAPK (ERK), and target of p70S6K1. PI3K and MAPK inhibitor, LY294002 and U0126 could inhibit hypoxia-induced HIF-1 and VEGF expression. We also investigated the role of reactive oxygen species (ROS) involved in HIF-1 and VEGF expression Exogenous addition of H2O2 was sufficient to activate Akt and ERK, scavengers of H2O2 significantly inhibited hypoxia-induced Akt and ERK, and subsequent HIF-lax expression and transcriptional activity. In conclusion, our data suggested that hypoxia- PI3K signaling through Akt and ERK kinases regulated ROS-dependent, hypoxia- induced HIF-1 activation and VEGF expression in gastric mucosa epithelial cells.

Liu L.;Ning X.;Han S.;Zhang H.;Sun L.;Shi Y.;Sun S.;Guo C.;Yin F.;Qiao T.;Wu K.;Fan D.

Molekuliarnaia Biologiia

2008

Hypoxia inducible factor-1α (HIF-1α) was well correlated with carcinogenesis and tumor progression in many kinds of cancer. In this study, high expression of HIF-1α was found in 37 of the 72 (51.39%) tumor specimens, and significantly correlated with venous invasion and lymphonode invasion. Patients with high expression of HIF-1α had a significantly shorter overall survival rate and disease-free survival rate than those with low expression. Multivariate analysis showed high HIF-1α expression was a borderline independent factor of overall survival. HIF-1α expression was also found to be significantly correlated with the expression of hepatitis B virus X protein (HBx), and over-expressed HBx upregulated HIF-1α protein expression in vitro. These results suggested that HIF-1α, which was partially regulated by HBx, might be a prognostic marker of HBV-related HCC patients. © 2008 Springer Science+Business Media, LLC.

Xie Huahong;Song Jiugang;Liu Kaige;Shen Huiqin;Hu Shengjuan;Yang Guitao;Du Yulei;Zou Xue;Jin Haifeng;Yan Li;Liu Jie;Fan Daiming;Ji Hongzan

Digestive Diseases and Sciences

2008

The present study aimed to describe the characterization of an antibody MGb2 that reacts with an epitope on gastric cancer cells, and identification of MGb2 antigen (MGb2-Ag). Immunostaining revealed its distribution in human tissues and demonstrated that the positive rate of MGb2-Ag was 81.48% in gastric cancer, 100% in gastric signet-ring cell carcinoma and mucinous adenocarcinoma, 13.16% in precancerous conditions, and 0% in chronic superficial gastritis. Using Western blotting, immunoprecipitation and MALDI-TOF MS (matrix assisted laser desorption/ionization time-of-flight mass spectrometry), MGb2-Ag was identified as TRAK1 (Trafficking protein, kinesin-binding 1), a new molecular gained limited recognition. Both MGb2 and commercial anti-TRAK1 Ab recognized prokaryotic expressed TRAK1. Immunostaining characteristics of TRAK1 were identical with MGb2-Ag in continuous sections of paraffin-embedded tissues of gastric tissues. This is the first report that TRAK1/MGb2-Ag is a promising diagnostic marker for gastric cancer and may help to detect signet-ring cell carcinoma and mucinous adenocarcinoma. © 2008 Elsevier Ireland Ltd. All rights reserved.

Zhang Faming;Ren Gui;Lu Yuanyuan;Jin Bin;Wang Jun;Chen Xiong;Liu Zhenxiong;Li Kai;Nie Yongzhan;Wang Xin;Fan Daiming

Cancer Letters

2009

Gastric cancer is the top lethal cancer in Asia. As the majority of cases present with advanced disease, conventional therapies (surgery, chemotherapy, and radiotherapy) have limited efficacy to reduce mortality. Emerging modalities provide promise to combat this malignancy. Target-protein-based cancer therapy has become available in clinical practice. Numerous molecules have been shown potential to target specific pathways for tumor cell growth. Cyclooxygenase-2 (COX-2) is overexpressed in and correlated with gastric cancer, and knockdown of COX-2 or administration of COX-2 inhibitors suppresses tumor formation in models of gastric cancer. Induction of apoptosis, reduction of angiogenesis, and blocking of potassium ion channels may present new mechanisms of COX-2 inhibition. Runt-related transcription factor 3 (RUNX3) is a candidate tumor suppressor gene whose deficiency is causally related to gastric cancer. RUNX3 is downregulated in metastatic gastric cancer. RUNX3 activation inhibits angiogenesis in xenograft tumors in nude mice. Tumor microenvironment modulation also provides a powerful tool to inhibit cancer development and progress; details of the potential roles of angiopoietins are discussed in this review. Osteopontin is a secreted protein involved in stress response, inflammation, wound healing, and immune response. Inhibition of osteopontin by RNA interfering technique suppressed tumorigenesis as well as angiogenesis in gastric cancer. Immunotherapy remains another important choice of adjuvant therapy for cancer. A tumor-specific antigen MG7-Ag has been identified with great potential for inducing immune response in gastric cancer. Using HLA-A-matched allogeneic gastric cancer cells to induce tumor-specific cytotoxic T lymphocytes appeared to be an alternative option of immunotherapy for gastric cancer. © 2009 The Authors.

Wu Kaichun;Nie Yongzhan;Guo Changcun;Chen Yu;Ding Jie;Fan Daiming

Journal of Gastroenterology and Hepatology Australia

2009

Li Xiaohua;Wu Kaichun;Fan Daiming

Hepatology

2009

We studied the subcellular localization of MAD2 in normal human tissues and gastric cancers. MAD2 showed nuclear and cytoplasmic localization in normal tissues such as muscle, testis, thyroid gland, cerebrum, trachea, and skin; blood vessels in some organs were also MAD2+. In normal stomach, MAD2 was expressed mainly in cytoplasm but showed nuclear staining in the majority of gastric cancers. MAD2 was significantly overexpressed in gastric cancer compared with matched adjacent tissues (P < .001), and expression was related to differentiation and other clinical parameters of cancer (P < .001). The cancer/adjacent normal tissue (C/N) ratio of MAD2 expression was higher than 2 and more frequently observed in patients with lymph gland metastasis (P < .05) and related to cancer differentiation. Our findings suggest that the steady-state amount of MAD2 inside cells may serve as a molecular switch in mitotic checkpoint control and that the subcellular localizations of this spindle protein undergo a shift during malignant transformation. The change of MAD2 expression may be involved mainly in gastric carcinogenesis and associated with the prognosis of gastric cancer; a C/N of more than 2 may be associated with the worse prognosis for survival in gastric carcinoma. © American Society for Clinical Pathology.

Wang Li;Yin Fang;Du Yulei;Du Wenqi;Chen Bei;Zhang Yongguo;Wu Kaichun;Ding Jie;Liu Jie;Fan Daiming

American Journal of Clinical Pathology

2009