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Objective To study the correlation between ABCC3 gene and bladder cancer cell proliferation,drug resistance and aerobic glycolysis.Methods Lipofectamine 2000 reagent was used for small interfering RNA (siRNA) transfection.Human ABCC3 siRNA and negative control siRNA were transfected into UMUC-3 and 5637 cells separately,and bladder cancer cells were divided into siABCC3-1 group,siABCC3-2 group and control group.Western blot assay was used to evaluate the ABCC3 expression levels.Quantitative real-time polymerase chain reaction (PCR) was performed to determine the ABCC3 mRNA expression levels.The effect of ABCC3 on bladder cancer cell growth was determined by colony formation assay.We also analyzed the sensitivity of cance cells to cisplatin by MTT assay.The effect of ABCC3 on aerobic glycolysis were detected by measuring LDHA protein levels,lactate production and glucose consumption.The measurement data were expressed as ((x) ± s) tandard deviation.The difference between the groups was analyzed by single factor analysis of variance and LSD.Results Both protein and mRNA levels of ABCC3 were significantly decreased after si-ABCC3 transfection.Bladder cancer cells treated with si-ABCC3 exhibited significantly lower colony numbers than that of the control group.5637 cells treated with siABCC3-1 and siABCC3-2 were 4.02 μg/ml and 3.91 μg/ml,respectively.The IC50 values of cisplatin after UMUC-3 cells siABCC3-1 and siABCC3-2 were 2.54 μg/ml and 2.49 μg/ml,respectively.The expression of lactate dehydrogenase A protein in bladder cancer cells was down-regulated and the expression of ABCC3 was positively correlated with the expression of LDHA (P =0.0362).siABCC3-1 and siABCC3-2 were transfected into 5637 cells,respectively.The glucose consumption decreased by 43.2％ and 43.7％ respectively.The lactic acid production was reduced by 31.3％ and 29.7％,respectively.After transfection of UMUC-3 cells with siABCC3-1 and siABCC3-2,glucose consumption decreased by 33.4％ and 37.5％,respectively,and lactic acid production decreased by 24.7％ and 25.2％,respectively,compared with the control group,the difference was statistically significant (P ＜ 0.001).Conclusions ABCC3 is an important oncoprotein involved in cell proliferation,glycolysis and drug resistance.It could be a promising predictive biomarker and potential therapeutic target for bladder cancer.
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BACKGROUND: Korea has high prescribing rate and rising antibiotic resistance. This study was conducted to understand why primary physicians prescribe antibiotics for acute respiratory infections and to explore the factors that influence antibiotic resistance, and so to suggest strategy to reduce antibiotic resistance. METHODS: A qualitative exploratory approach was used using 4 focus groups composed of physicians from different area. A semi-structured guide was applied in obtaining the physicians' opinions. Common themes were extracted by authors, which were used to gather results and draw conclusion. RESULTS: Participants acknowledged multiple factors such as clinical factor and competitive environment are involved in physicians' decision of antibiotic prescribing. They identified that causes of rising antibiotic resistance were shortage of information, discontinuation of taking antibiotics, and other system factors. CONCLUSION: Participants were certain that less prescribing antibiotics and selecting appropriate antibiotics might be method to reduce antibiotic resistance. To change the prescribing behavior, it should be provided periodically for community physicians with prescribing information and specific guidelines for antibiotics resistance. Patients should be also noticed about antibiotic medication information more accurately. Including prescription incentive policy, improvement of healthcare system will be carried out at the same time.
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Humans , Anti-Bacterial Agents , Delivery of Health Care , Drug Resistance, Microbial , Focus Groups , Information Dissemination , Korea , Methods , Motivation , Prescriptions , Respiratory Tract InfectionsABSTRACT
Objective To investigate clinical features and antibiotic resistance of pseudomonas aeruginosa (PA) strains isolated from patients with diabetic foot infections (DFI) in Tianjin Metabolic Diseases Hospital.Methods Eighty-five PA strains were isolated from 428 patients with diabetic foot in the hospital from Jan 2008 to Dec 2010.The clinical features of patients were summarized.Relationships between the isolates and depth of ulcer or severity of infection were analyzed.The disk-diffusion method was performed to examine antimicrobial susceptibility.Results Gram positive (G+) and Gram negative (Gˉ) isolates were 50.47％ and 41.12％,respectively.Multidrug-resistant PA composed 32.9％ of the total PA isolates.The size of ulcers with PA infections was bigger than those with non-PA bacterial infections (P＜0.05).Compared to G+ strains,patients with PA strains were older,had lower hemoglobin,but higher serum sensitive C-reactive protein; and more frequently,they had ischemic ulcer and osteomyelitis.Compared to G+ strains,the PA strains were more frequently isolated from deeper ulcers and with more serious infections(P＜0.05).The resistant rates of PA to cephalosporins,fluoroquinolones,and aminoglycosides were between 32.9％-61.2％,37.6％-42.4％,and 37.6％-62.4％,respectively.Only one out of 85 PA strains was imipenem-resistant.However,sensitiveness of all PA isolates to cefoperazone and sulbactam reached 100％.Conclusion PA strains are mainly found in patients with deeper ulcers and more serious infections.Multidrug-resistant PA is common in DFI.It is important to isolate pathogens and determine their antibiotic resistance correctly in diabetic foot patients in order to provide appropriate drug administration and to reduce the production and dissemination of drug resistant strains.
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It has been preliminarily demonstrated that CIAPIN1 plays an essential role in normal tissue and is closely related to tumor formatin and development, prognosis and multiple drug resistance. CIAPIN1-based target therapy may provide a novel thinkingpathway in cancer treatment.
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Background and purpose:Multidrug resistance(MDR)is the main obstacle of chemotherapy in the treatment of cancer,and it has been reported that the muted p53 gene is related to MDR.In this study,we explored whether adenovirus mediated p53 gene(Ad-p53)could reverse MDR of human breast cancer and its impacts on the expressions of P-gp,TOPOⅡ and GST-?.Methods:In this study,adriamycin-resistant human breast carcinoma cells(MCF-7/Adr)and its parental cells(MCF-7)were used to determine the effect of Ad-p53.Cck-8 assay was adopted to evaluate the cytotoxicity of adriamycin.Western blot were performed to observe the expression of P-glycoprotein(P-gp),TopoismeraseⅡ(TOPOⅡ)and GST-?.Results:After transfection with 50 MOI Ad-p53,the 50% inhibitory concentration(IC50)of adriamycin to MCF-7/Adr cells was decreased from(4.54?0.91)?g/ml to(0.26?0.11)?g/ml,and the chemosensitivity increased 18.1 times(P
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Objective To establish the drug resistant cell line of choriocarcinoma and to study the transfection of the human interleukin 2 (hIL 2) gene into the established drug resistant cell line and investigate the reversal of the multidrug resistance Methods The resistant cell line was established by pulse exposed choriocarcinoma cell line JEG 3 to etopside (VP 16) for ten months The recombinant plasmid containing pcDNA3 1(+) hIL 2 gene was constructed The drug resistant cell line was transfected with the constructed plasmid by lipofectin, and the tumor cell colonies containing the IL 2 sequence were selected by genetin The expression of hIL 2 and drug resistant related genes was detected by reverse transcript polymerase chain reaction The chemosensitivity of the gene transfected tumor cells and the non transfected cell lines to methetraxate, VP 16, kengshengmycine, paclitaxol and 5 fluorouracil was determined by the methyl thiazolyl tetrazolium cytotoxicity assay Results The transfected cells expressed human hIL 2 gene, and showed the reversal of multidrug resistance by methyl thiazolyl tetrazolium assay The transfected cells expressed no multidrug resistance gene 1 (MDR1) on mRNA level Drug resistance index to VP 16 decreased from 38 7 to 6 0 and 6 1, the index to methetraxate decreased from 14 5 to 2 6 and 2 5, to methetraxate from 13 0 to 2 0 Conclusion The transfection of hIL 2 gene into the drug resistance cell line of choriocarcinoma can modulate the MDR1 expression on the mRNA level, and reverse the drug resistance