Objective To explore the role of Vav3 gene in multidrug resistance (MDR) of gastric cancer and the related mechanism. Methods QRT-PCR was used to examine the expressions of Vav3 gene in gastric cancer tissues, tumor-adjacent tissues, human gastric cancer cell line SGC7901, and gastric epithelial cell line GES-1. Then Vav3-siRNA was synthesized and tansfected into SGC7901 cells. MTT assay was then used to determine the inhibition rates of tumor cells exposed to chemotherapeutic agents (5-FU, L-OHP) before and after Vav 3-siRNA transfection. Realtime RT-PCR and Western blotting analysis were used to observe the expressions of inhibitor of apoptosis proteins (IAPs) : xIAP, Survivin, and Livin; meanwhile, the expression and activity of Caspase-3 and Caspase-8 were also determined. Results Vav3 was over-expressed in gastric cancer tissues and gastric cell line compared with those in tumo-adjacent tissues and gastric epithelial cell line GES-1 (P<0.05). Expression of Vav3 was significantly inhibited by Vav3-siRNA (P<0.01). Inhibition rates of tumor cells exposed to 5-FU and L-OHP were significantly increased 48 h after Vav3-siRNA tansfection (P<0.05). The expressions of xIAP and Survivin were significantly decreased in cancer cells after Vav3-siRNA tansfection (both P<0.05), and no notable change was found for Livin expression; also the expression and activity of Caspase-3 and Caspase-8 protein were significantly increased after Vav3-siRNA tansfection in SGC7901 cells (all P<0.05). Conclusion Vav3 can participate in MDR of gastric cancer by regulating apoptotic pathways, and inhibition of Vav3 can help reverse MDR of gastric cancer cells by regulating some IAPs.

0.05).Conclusions The higher level of SEB-specific IgM and IgE in AD and eczema indi cates the colonization of Staphylococcus aureus,which participates in the exace rbation of allergic inflammation,is involved in the pathogenesis of AD and ecz ema.

Objective To analysis the etiology and molecular classification of brucella strains isolated in Guangdong province in 2010.Methods The strains of 19 brucella were verified and identified by some methods including traditional biology phenotype confirmation,PCR amplification and pulsed field gel electrophoresis (PFGE).Results On phenotype level,4 strains were brucella melitensis biovar 1,2 strains were brucella suis biovar 3,and the rest were brucella melitensis biovar 3,which were specific B genes positive strains,and the PFGE typing similar values ranging from 67.9％ to 100％.In addition to the four strains from Zhuhai for the outbreak,the homology was 100％,and the rest were sporadic cases.Conclusions Brucella cases,in Guangdong province,are highly sporadic and dispersed outbreaks.Compared with a few years ago,it shows species diversification,and brucella melitensis biovar 3 is still the dominant serotype.PFGE can be used to distinguish the three species of brucella,but it can't effectively distinguish the allotypes.

BACKGROUNDIntegrase interactor 1 (INI1), which encodes a component of the ATP-dependent chromatin remodeling hSWI-SNF complex, has been identified as a tumor suppressor in many tumors. Nonetheless, the role of INI1 in gastric tumor progression is not known exactly. The aim of this research was to investigate the effect of INI1 in the carcinogenesis and progression of gastric cancer.

METHODSGastric tumor tissues with different differentiation levels from clinical gastric carcinoma samples and adjacent control normal tissues were taken. Expression levels of INI1 were detected by quantitative reverse transcriptation-polymerase chain reaction (RT-PCR) and Western blotting. Gastric cancer cell line SGC7901 was transfected with INI1 eukaryotic expressing vector INI1-GFP. Cell proliferation activities were assessed by MTT; cell count and cell cycle were detected by flow cytometry (FCM); cell apoptosis were measured by TUNEL and FCM; cell migration and invasiveness were evaluated by wound healing and transwell assays. Expression levels of INI1 and proliferation-related genes including p16, p21, cyclin D1 and cyclin A, apoptosis genes p53, B-cell non-Hodgkin lymphoma-2 (Bcl-2), Bcl-2-associated x protein (Bax) and caspase-3, and invasion-related genes including intercellular adhesion molecule 1 (ICAM1), matrix metalloproteinase 2 (MMP2), MMP9 and tissue inhibitor of matrix metalloproteinase 1 (TIMP1), were detected by quantitative RT-PCR and Western blotting.

RESULTSINI1 expression levels were lower in gastric carcinoma compared with adjacent control normal tissues. Overexpression of INI1 in SGC7901 cells inhibited its proliferation and invasiveness, but increased anoikis and G(0)/G(1) cell number. INI1-GFP transfection upregulated expression of INI1 and proliferation related genes p16 and p21, apoptosis genes p53 and Bax, and invasion-related genes TIMP1; cyclin D1, cyclin A, Bcl2, ICAM1, MMP2 and MMP9 were downregulated, and there was no significant change in caspase 3 levels.

CONCLUSIONINI1 plays a key role in gastric carcinogenesis by affecting proliferation, apoptosis and invasion.

Apoptosis ; genetics ; physiology ; Blotting, Western ; Cell Cycle ; genetics ; physiology ; Cell Line, Tumor ; Cell Proliferation ; Chromosomal Proteins, Non-Histone ; genetics ; metabolism ; DNA-Binding Proteins ; genetics ; metabolism ; Humans ; Real-Time Polymerase Chain Reaction ; SMARCB1 Protein ; Stomach Neoplasms ; genetics ; metabolism ; Transcription Factors ; genetics ; metabolism

OBJECTIVETo investigate the differentiation-related proteins in human gastric carcinoma cell lines by comparative proteomics.

METHODSThe holoproteins of human gastric carcinoma cell lines MKN28, SGC7901 and BGC823 were measured by two-dimensional gel electrophoresis and matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Some proteins identified by proteomics were tested by Western blot in the cell strains and tissues of gastric carcinoma.

RESULTS14 differential protein spots were found in the 3 gastric carcinoma cell lines, among them 8 spots were identified by MALDI-TOF-MS. These proteins were probably thioredoxin peroxidase, glyceraldehyde-3-phosphate dehydrogenase (GAPD), beta-tubulin polypeptide, hypothetical protein, zinc finger protein (ZNF) 139, protein-tyrosine kinase, calreticulin precursor, and tropomyosin, proteins related with biological behavior of gastric carcinoma cells such as signal transduction, cellular homeostasis, glycolysis, antioxidation action, multidrug resistance(MDR), etc. The expressions of those proteins in gastric cancer cells and tissues identified by Western blot were consistent with the results obtained by proteomics.

CONCLUSIONDifferential proteins are found in 3 human gastric carcinoma cell lines, mainly, proteins related with cell signaling, maintenance of homeostasis, glycolysis, metabolism of anti-cancer drug and anti-oxidative injury, etc.

Adenocarcinoma ; metabolism ; pathology ; Adult ; Aged ; Blotting, Western ; Cell Line, Tumor ; Electrophoresis, Gel, Two-Dimensional ; Female ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Glyceraldehyde-3-Phosphate Dehydrogenases ; metabolism ; Humans ; Male ; Middle Aged ; Protein-Tyrosine Kinases ; metabolism ; Proteome ; analysis ; Proteomics ; methods ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Stomach Neoplasms ; metabolism ; pathology

OBJECTIVETo explore the effects of antisense oligodeoxynucleotides (ASODN) on proliferation and apoptosis in gastric cancer cell line BGC-823 cells and the molecular mechanisms induced by ASODN.

METHODSsurvivin ASODN-1, survivin ASODN-2 and survivin ASODN-3 were transfected into BGC-823 cells by Lipofectamine(TM) 2000 transfection reagent. The growth activity of BGC-823 cells was detected by MTT assay. Apoptosis index (AI), proliferation index (PI), cell cycle and expressions of survivin, VEGF and Smac/DIABLO proteins were detected by flow cytometry (FCM). The changes of survivin mRNA, VEGF mRNA and Smac/DIABLO mRNA were detected by RT-PCR.

RESULTSThe expression of survivin was down-regulated by the three ASODN sequences, especially the ASODN-2 was best. At 48 hours after transfection with 600 nmol/L survivin ASODN-2, the cells in G(1)/G(0) phase were significantly increased [(72.25 ± 2.95)%], apoptotic index increased [(11.31 ± 0.38)%], proliferation index decreased [(27.77 ± 2.97)%], compared with those in the control group [(56.25 ± 0.75)%, (1.62 ± 0.36)%, (43.80 ± 0.80)%, all P < 0.05]. The survivin mRNA and protein levels (0.523 ± 0.091, 0.733 ± 0.009) were down-regulated compared with those in the control group (0.861 ± 0.047, 0.997 ± 0.233), VEGF (0.519 ± 0.076, 0.75 ± 0.006) were down-regulated compared with those in the control group (0.779 ± 0.059, 1.000 ± 0.01), while those of Smac/DIABLO(0.899 ± 0.113, 1.637 ± 0.023)were up-regulated compared with those in the control group (0.558 ± 0.041, 1.000 ± 0.049, all P < 0.05).

CONCLUSIONSSurvivin ASODN can induce apoptosis and inhibit the proliferation of gastric cancer cell line BGC-823 cells. Those effects are induced through up-regulation of Smac/DIABLO and down-regulation of survivin and VEGF expression simultaneously.

Apoptosis ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Down-Regulation ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; metabolism ; Intracellular Signaling Peptides and Proteins ; genetics ; metabolism ; Mitochondrial Proteins ; genetics ; metabolism ; Oligodeoxyribonucleotides, Antisense ; genetics ; RNA, Messenger ; metabolism ; Stomach Neoplasms ; genetics ; metabolism ; pathology ; Transfection ; Up-Regulation ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

OBJECTIVETo investigate the expression features of P-glycoprotein (P-gp), glutathione S transferase-pi (GST-pi) and inhibitor of apoptosis proteins like p53, survivin and bcl-2 in lymph node metastases of gastrointestinal carcinomas.

METHODSThe expression of P-gp, GST-pi, p53, survivin and bcl-2 were determined by using immunohistochemistry technique in surgical specimens of primary tumor (PT) and lymph node metastases (LNMs) from 54 gastrointestinal cancer patients with metastasis of lymph nodes. The expression difference of 5 multi-drug resistance (MDR)-related factors between LNMs and PT were compared.

RESULTSSignificant difference was found in the expression of P-gp and GST-pi between the two groups (both P < 0.05), and expression of p53 and bcl-2 showed positive correlation between LNMs and PT (r = 0.7248, 0.5524; both P < 0.05), respectively. In LNMs, P-gp expression was positively correlated with GST-pi (r = 0.4062, P < 0.05) and survivin (r = 0.6169, P < 0.05), and also GST-pi expression was related positively with survivin (r = 0.4027, P < 0.05). Statistically positive correlations were noted between bcl-2 and P-gp (r = 0.3986, P < 0.05), bcl-2 and survivin (r = 0.2937, P < 0.05), as well as GST-pi and survivin (r = 0.4481, P < 0.01) in PT. Only a positive correlation between GST-pi and survivin expression was simultaneously shown in both LNMs and PT.

CONCLUSIONSThere is significant heterogeneity of MDR-related factors expression in LNMs of gastrointestinal carcinomas. Effective adjuvant chemotherapy after operation should target on the metastatic loci of the disease.

ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Adult ; Aged ; Aged, 80 and over ; Digestive System Neoplasms ; metabolism ; pathology ; Female ; Glutathione S-Transferase pi ; metabolism ; Humans ; Inhibitor of Apoptosis Proteins ; Lymph Nodes ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Microtubule-Associated Proteins ; metabolism ; Middle Aged ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Tumor Suppressor Protein p53 ; metabolism

Objective To investigate the effects of different dialysates on expression of protein kinase C-? (PKC?) and apoptosis of U937 cell line. Methods Different dialysates were added into culture fluid with U937 cell line at exponential phase of growth, and groups were divided: fluid A+fluid B group (dialysate A+dialysate B), fluid A+fluid B+rottlerin (PKC? specific inhibitor)group, fluid A+powder B group (dialysate A+powder B) and fluid A+powder B + rottlerin group. Besides, blank control group and normal control group were established. Cells were harvested 24 h and 48 h after treatment, morphological changes were observed by Hoechst33258 fluorescence staining, cell apoptosis was measured by Annexin-V-FITC/PI double staining, and expression of PKC? mRNA and protein was detected by RT-PCR and Western blotting, respectively. Results Cell apoptosis significantly increased in fluid A+powder B group, with typical morphology of apoptosis. After treatment for 24 h and 48 h, cell apoptosis rates in fluid A+powder B group were significantly higher than those at corresponding time points in blank control group, normal control group and fluid A+powder B+rottlerin group (P0.05). Conclusion Fluid A+powder B can significantly increase apoptosis of U937 cell line, the mechanism of which may be associated with the up-regulation of expression of PKC?. Compared with fluid A+powder B, fluid A+fluid B is superior in reducing apoptosis of peripheral blood monouclear cells.

Objective To analyze the etiologic characteristics of Vibrio cholerae in Guangdong province in 2007.Genetic relationship was observed including among predominated biotype isolates from different areas within the province and among same biotypes isolates from cholera cases and regular surveillance.Methods Isolates from cholera cases and through environmental surveillance were typed by sero-and phage-typings.Similarity of molecular fingerprinting was analyzed through comparing the pulsed field gel electrophoresis(PFGE)pattern of predominated biotype isolates,and those of the same biotype isolates from cholera cases and environment surveillance,respectively.In addition,genetic relationship was determined by clustering analysis,using bionumerics software.Results In total,31 isolates from cholera cases were collected and subtyped for 3 serogroups.V.cholerae O1 El Tor Inaba phage 1d was the predominant biotype which causing most of the cases in Guangdong province in 2007.Data from cluster analysis showed that the similarity among Inaba phage 1d strains from different areas were from 94.5% to 100%.However.16 isolates were collected from environment surveillance programs and the predominated biotype could not be found.Additionally,the biotype distribution of cases isolates was not consistent with those isolates through surveillance.High phylogenetic diversity was observed for the same biotypes isolates from cases and surveillance samples.Conclusion Our data showed that V.cholerae O1 El Tor Inaba phage 1d was the predominated biotype with multi-clone coexisting and circulating in Guangdong province in 2007.It also appeared to be the characteristics of cholera in the non-epidemic period,suggesting that it was necessary to enhance the alert surveillance programs for cholera epidemic based on the molecular typing techniques.