CLIC1 knockout inhibits invasion and migration of gastric cancer by upregulating AMOT-p130 expression

Purpose To explore the regulatory relationship between Chloride intracellular channel 1 (CLIC1) and Angiomotin (AMOT)-p130, and reveal the role of AMOT-p130 in gastric cancer (GC). Methods Immunohistochemistry was performed to analyze the expression of CLIC1 and AMOT-p130 in GC tissues and adjacent tissues. The expression of AMOT-p130 upon CLIC1 silencing was analyzed using RT-PCR, western blot, and immunofluorescence in GC cells. Transwell and wound-healing assays were performed to detect migration and invasion in GC cells. The changes in EMT-related proteins were detected using western blot. Results Our study found that high CLIC1 expression was significantly associated with low AMOT-p130 expression in GC tissues. Silencing CLIC1 expression in MGC-803 cells (MGC-803 CLIC1 KO) and AGS cells (AGS CLIC1 KO) decreased the invasive and migratory abilities of tumor cells, which were induced by the upregulation of AMOT-p130. Subsequently, we demonstrated that AMOT-p130 inhibits the invasive and migratory abilities of GC cells by inhibiting epithelial–mesenchymal transition. Conclusions Our study suggests that AMOT-p130 could inhibit epithelial–mesenchymal transition in GC cells. CLIC1 may participate in the metastatic progression of GC by downregulating the expression of AMOT-p130 (AU)

CLIC1 knockout inhibits invasion and migration of gastric cancer by upregulating AMOT-p130 expression.

PURPOSE: To explore the regulatory relationship between Chloride intracellular channel 1 (CLIC1) and Angiomotin (AMOT)-p130, and reveal the role of AMOT-p130 in gastric cancer (GC). METHODS: Immunohistochemistry was performed to analyze the expression of CLIC1 and AMOT-p130 in GC tissues and adjacent tissues. The expression of AMOT-p130 upon CLIC1 silencing was analyzed using RT-PCR, western blot, and immunofluorescence in GC cells. Transwell and wound-healing assays were performed to detect migration and invasion in GC cells. The changes in EMT-related proteins were detected using western blot. RESULTS: Our study found that high CLIC1 expression was significantly associated with low AMOT-p130 expression in GC tissues. Silencing CLIC1 expression in MGC-803 cells (MGC-803 CLIC1 KO) and AGS cells (AGS CLIC1 KO) decreased the invasive and migratory abilities of tumor cells, which were induced by the upregulation of AMOT-p130. Subsequently, we demonstrated that AMOT-p130 inhibits the invasive and migratory abilities of GC cells by inhibiting epithelial-mesenchymal transition. CONCLUSIONS: Our study suggests that AMOT-p130 could inhibit epithelial-mesenchymal transition in GC cells. CLIC1 may participate in the metastatic progression of GC by downregulating the expression of AMOT-p130.

Effects of miR-153 on angiogenesis in MCAO rats through Shh signaling pathway.

OBJECTIVE: Cerebrovascular disease is a disease which has the highest mortality in China. Angiogenesis in the ischemic region after cerebral infarction is closely related to its prognosis. Recent studies found that microRNAs (miRNAs) are involved in the regulation of neovascularization. MicroRNA-153 (MiR-153) has protective effects on the ischemic injury, but its relationship with the Sonic Hedgehog (Shh) signaling pathway is still unclear. This work aimed to investigate the role of miR-153 in angiogenesis of middle cerebral artery occlusion (MCAO) rats through the Shh signaling pathway. MATERIALS AND METHODS: The rat cerebral ischemic injury (MCAO) model was established by thread embolism and treated by Agomir-153 and 5-EI. MiR-153 expression was detected using Real Time-Polymerase Chain Reaction (RT-PCR). The neurological function was assessed. The infarct area of the brain and the capillary density were determined using 2,3,5-triphenyl tetrazolium chloride (TTC) method. The Shh signaling pathway and angiogenesis-related factors were tested by Western blot assay. RESULTS: Agomir-153 or Agomir-153 combined with 5-EI significantly increased miR-153 expression, reduced the infarct area, and promoted the generation of cerebral capillaries in the MCAO model. 5-EI partially blocked the protective effects of Agomir-153 and angiogenesis. The up-regulation of miR-153 markedly inhibited patched (PTC) expression and activated the Shh signaling pathway. CONCLUSIONS: The up-regulation of miR-153 rats activated the Shh signaling pathway to promote angiogenesis and improve prognosis through lipid-coated Patch (PTC) in MCAO. MiR-153 was considered to be a new therapeutic target for promoting angiogenesis after MCAO.

Rupture of liver abscess following hepatogastric fistula caused by perforation of remnant gastric carcinoma: a case report.

OBJECTIVE: We report the case of a 73-year-old man, with a history of proximal subtotal gastrectomy, who suffered acute abdominal symptoms and signs. Laparotomy showed rupture of liver abscess and hepatogastric fistula formation caused by perforation of remnant stomach. CASE REPORT: Residual stomach resection, incision and drainage of liver abscess were performed, and the patient was smoothly discharged from hospital nineteen days after the emergency operation. RESULTS: The final pathology confirmed the remnant gastric adenocarcinoma. This case is so far the first reported liver abscess caused by perforation of residual stomach malignant tumor. CONCLUSIONS: Liver abscess and hepatogastric fistula are rare. This is the first report on a remnant gastric adenocarcinoma (RGC) invading the adjacent liver, with ruptured liver abscess resulting from gastric perforation. We speculated that there were inevitable factors for this case. Direct invasion to the liver capsule of gastric carcinoma was the bridging basic of the formation of a hepatogastric fistula. Pyloric obstruction caused by gastric carcinoma was the driver of liver abscess rupture since the increased proximal gastrointestinal pressure led to the inner pressure of liver abscess rising through the conduction of hepatogastric fistula. The recommended treatment protocol for this clinical entity comprises removal of the primary lesions and drainage of the liver abscess. This successful case provided us with a great deal of clinical information and treatment experience.

Impact of bilateral subcostal rooftop incision combined double lifting retractor on recent surgical outcomes of obese patients with advanced gastric cancer.

OBJECTIVE: To investigate the impact of bilateral subcostal rooftop incision under bilateral costal margins combined double lifting retractors on the recent surgical outcome of obese patients with advanced gastric cancer. PATIENTS AND METHODS: We retrospectively analyzed the clinical data of 138 obese patients with gastric cancer under radical gastrectomy. The patients were divided into two groups, the MI group (n=68) and RI group (n=70). Patients in MI group were treated by midline incision via upper abdomen combined single double lifting retractor technology while the patients in RI group were treated by roof incision under bilateral costal margins under combined double lifting technology. The abdomen entering and closing time, the operation time, bleeding volume, blood transfusion rate, postoperative drainage volume, postoperative hospitalization time, postoperative detected number of perigastric lymph node, postoperative pain score, postoperative complications and short-term survival between the two groups were compared and analyzed. RESULTS: The average abdominal entering and closing time of RI group were slightly longer than MI group, but the average total operation time of RI group was shorter than MI group. The average bleeding volume was 98.4 ± 46.8 ml, the postoperative intraperitoneal drainage was 24 h (123.8 ± 69.0 ml), 48 h (101.9±36.7 ml), 72 h (45.1±24.6 ml), and the postoperative hospitalization time was 12.9 ± 2.5 d, and the incidence of postoperative complications was 17.1%, all of which were less than MI group. The difference on the average total detected number of perigastric lymph node between the two groups was not statistically significant. 129 cases (93.5%) were followed up for 12 to 36 months, and the difference on survival curve between the two groups was not statistically significant (p>0.05). CONCLUSIONS: Bilateral subcostal rooftop incision was favorable for exposing the stomach, adjacent organs and large vessels, convenient for operation and would not affect the postoperative recovery, so it was a good choice for obese patients with gastric cancer.

Survivin as a therapeutic target in Sonic hedgehog-driven medulloblastoma.

Medulloblastoma (MB) is a highly malignant brain tumor that occurs primarily in children. Although surgery, radiation and high-dose chemotherapy have led to increased survival, many MB patients still die from their disease, and patients who survive suffer severe long-term side effects as a consequence of treatment. Thus, more effective and less toxic therapies for MB are critically important. Development of such therapies depends in part on identification of genes that are necessary for growth and survival of tumor cells. Survivin is an inhibitor of apoptosis protein that regulates cell cycle progression and resistance to apoptosis, is frequently expressed in human MB and when expressed at high levels predicts poor clinical outcome. Therefore, we hypothesized that Survivin may have a critical role in growth and survival of MB cells and that targeting it may enhance MB therapy. Here we show that Survivin is overexpressed in tumors from patched (Ptch) mutant mice, a model of Sonic hedgehog (SHH)-driven MB. Genetic deletion of survivin in Ptch mutant tumor cells significantly inhibits proliferation and causes cell cycle arrest. Treatment with small-molecule antagonists of Survivin impairs proliferation and survival of both murine and human MB cells. Finally, Survivin antagonists impede growth of MB cells in vivo. These studies highlight the importance of Survivin in SHH-driven MB, and suggest that it may represent a novel therapeutic target in patients with this disease.

Biomass derived furfural-based facile synthesis of protected (2S)-phenyl-3-piperidone, a common intermediate for many drugs.

An efficient synthetic route towards tosyl-protected (2S)-phenyl-3-piperidone, a common intermediate for many drugs, has been developed in 5 steps in 54% yield from biomass derived furfural. The synthetic utility of the piperidone core structure was demonstrated with the synthesis of a NK1 receptor antagonist.

Identification of biomarkers for lymph node metastasis in early-stage cervical cancer by tissue-based proteomics.

BACKGROUND: Pelvic lymph node metastasis (PLNM) is the key to determining the treatment and prognosis of early-stage cervical cancer (CC, I-IIst). The aim of this study was to identify biomarkers for PLNM of CC, I-IIst. METHODS: Two-dimensional fluorescence difference gel electrophoresis and matrix-assisted laser desorption/ionisation-time-of-flight mass spectrometry (MALDI-TOF/TOF MS) were used to identify differentially expressed proteins in primary CC, I-IIst tissue with (n=8) and without (n=10) PLNM. The expression levels of three differential proteins (FABP5, HspB1, and MnSOD) were validated using western blotting and immunohistochemistry. An independent cohort of 105 CC, I-IIst patients was analysed to assess the correlation of FABP5, HspB1, and MnSOD with clinicopathologic factors and clinical outcomes. RESULTS: Forty-one differential proteins were identified. Upregulation of FABP5, HspB1, and MnSOD in CC, I-IIst with PLNM was confirmed and was significantly correlated with PLNM. FABP5, HspB1, and MnSOD were significant predictors of PLNM in univariate analysis. FABP5, HspB1, and lymphovascular space invasion (LVSI) were independent predictors of PLNM in multivariate analysis. Survival curves indicated that CC, I-IIst patients with FABP5, HspB1, and MnSOD upregulation had poor prognosis. CONCLUSIONS: FABP5, HspB1, and MnSOD may be potential biomarkers for PLNM of CC, I-IIst and may have important roles in the pathogenesis of PLNM.

YAP modifies cancer cell sensitivity to EGFR and survivin inhibitors and is negatively regulated by the non-receptor type protein tyrosine phosphatase 14.

The Yes-associated protein (YAP) is a transcriptional factor involved in tissue development and tumorigenesis. Although YAP has been recognized as a key element of the Hippo signaling pathway, the mechanisms that regulate YAP activities remain to be fully characterized. In this study, we demonstrate that the non-receptor type protein tyrosine phosphatase 14 (PTPN14) functions as a negative regulator of YAP. We show that YAP forms a protein complex with PTPN14 through the WW domains of YAP and the PPXY motifs of PTPN14. In addition, PTPN14 inhibits YAP-mediated transcriptional activities. Knockdown of YAP sensitizes cancer cells to various anti-cancer agents, such as cisplatin, the EGFR tyrosine kinase inhibitor erlotinib and the small-molecule antagonist of survivin, S12. YAP-targeted modalities may be used in combination with other cancer drugs to achieve maximal therapeutic effects.

The safety of Bacillus subtilis and Bacillus indicus as food probiotics.

AIMS: To conduct in vitro and in vivo assessments of the safety of two species of Bacillus, one of which, Bacillus subtilis, is in current use as a food supplement. METHODS AND RESULTS: Cultured cell lines, Caco-2, HEp-2 and the mucus-producing HT29-16E cell line, were used to evaluate adhesion, invasion and cytotoxicity. The Natto strain of B. subtilis was shown to be able to invade and lyse cells. Neither species was able to adhere significantly to any cell line. The Natto strain was also shown to form biofilms. No strain produced any of the known Bacillus enterotoxins. Disc-diffusion assays using a panel of antibiotics listed by the European Food Safety Authority (EFSA) showed that only Bacillus indicus carried resistance to clindamycin at a level above the minimum inhibitory concentration breakpoints set by the EFSA. In vivo assessments of acute and chronic dosing in guinea pigs and rabbits were made. No toxicity was observed in animals under these conditions. CONCLUSIONS: Bacillus indicus and B. subtilis should be considered safe for oral use although the resistance of B. indicus to clindamycin requires further study. SIGNIFICANCE AND IMPACT OF THE STUDY: The results support the use of B. subtilis and B. indicus strains as food supplements.

Aldol reaction under solvent-free conditions: highly stereoselective synthesis of 1,3-amino alcohols

[formula: see text] A method for the highly stereoselective synthesis of 1,3-amino alcohols based on the indium trichloride-catalyzed Mukaiyama aldol reaction of keto ester (R,S)-1 under solvent-free conditions has been developed. The high selectivity observed can be explained on the basis of the shielding of one face by a remote substituent.