A novel β1,3-N-acetylglucosaminyltransferase involved in invasion of cancer cells as assayed in vitro
45
Citation
29
Reference
10
Related Paper
Citation Trend
目的研究极低频电磁场暴露对小鼠脑和肝组织c-fos mRNA水平的影响.方法将小鼠暴露于50 Hz、0.2 mT及50 Hz、6.0 mT的电磁场中,持续2周或4周;采用竞争性RT-PCR方法检测小鼠脑和肝脏c-fos mRNA水平.结果50 Hz、0.2 mT及50 Hz、6.0 mT电磁场暴露2周后,小鼠脑组织c-fos mRNA的水平上升为(0.017 8±0.007 6)amol/120 ng cDNA和(0.009 2±0.004 2)amol/120 ng cDNA,与对照组[(0.001 2±0.000 5)amol/120 ng cDNA]比较,差异有显著性(P＜0.05);肝组织c-fos mRNA的水平上升为(0.0117±0.005 5)amol/120ng cDNA和(0.014 8±0.016 2)amol/120 ng cDNA,与对照组[(0.000 5±0.000 5)amol/120 ng cDNA]比较,差异有显著性(P＜0.05).0.2 mT和6.0 mT电磁场暴露4周后,小鼠脑组织c-fos mRNA的水平上升为(0.010 0±0.005 4)amol/120 ng cDNA和(0.019 8±0.007 9)amol/120 ng cDNA,与对照组[(0.001 5±0.000 8)amol/120ng cDNA]比较,差异有显著性(P＜0.05);肝组织c-fos mRNA的水平分别上升为(0.017 3±0.012 2)amol/120ng cDNA和(0.013 3±0.009 0)amol/120 ng cDNA,而对照组无表达.结论50 Hz电磁场暴露引起小鼠脑和肝脏c-fos基因转录水平明显上调.
Cite
Citations (0)
Cellular transfections provide powerful experimental tools to study gene regulation. However, endothelial cells are difficult to transfect. Therefore, the present studies were performed to optimize transfection techniques in human endothelial cells. To study transfection rates, endothelial cells were transfected with the pGL3 vector, containing the luciferase reporter gene, complexed with several currently available liposomes, such as different Perfect Lipid (pFx) mixtures, DMRIE‐C, or lipofectin. The optimal transfection rate was achieved in cells transfected for 1.5 h with 5 mg/mL of DNA plasmid in the presence of 36 mg/mL of pFx‐7. Transfections mediated by other liposomes were less efficient. Usefulness of the optimized transfection technique was confirmed in cells transfected with NF‐kB or AP‐1‐responsive constructs. We conclude that among several currently available liposomes, pFx‐7 appears to be the most suitable for transfections of cultured human endothelial cells. Acknowledgements: Supported by NIH; NS39254, MH63022, and AA013843.
Cite
Citations (4)
The study aimed at improving gene transfection efficiency. We used PEI-mediated plasmid pEGFP-N1 to transfect NIH-3T3 cells in this study. NIH-3T3 Cells were divided into two groups to conduct one and two transfection,and observed under fluorescent inverted microscope at 48 h after transfection. The results showed that some cells emited green fluorescence,and green fluorescent cells in two transfection group were more than one transfection group. Calculated transfection efficiency to showed that two transfection method could significantly improve gene transfection efficiency.
3T3 cells
Cite
Citations (0)
Abstract We constructed a “cDNA bank” of human colorectal cancer and surrounding normal tissues with our unique mRNA assay system. Total nucleic acids extracted from patients’ tissues were applied to 96-well plates, where poly(dT) sequences of oligonucleotides were immobilized. After hybridization, the cDNA was reverse-transcribed on the plate with the captured mRNA as a template, followed by synthesis of double-stranded (ds) cDNA. The resulting sense cDNA was removed from the plate, then used in PCR for analysis of various genes. The sense strand of the cDNA was repeatedly synthesized by using the immobilized antisense cDNA as a template even from plates used once and stored at 4 °C for as long as 6 months. Furthermore, the results of PCR could be easily compared among different specimens if the same amount of total mRNA were applied to the plate for the ds cDNA synthesis. This demonstrated that the cDNA bank constructed from clinical materials provides almost unlimited supplies of cDNA for multiple gene analysis of cancer.
Cite
Citations (4)
Objective: To evaluate the transfection efficiency and optimal conditions of siRNA.Methods:Synthysized siRNA were transfected into human umbilical vein endothelial cells,we identified the rate of the transfection by fluorescent microscope and Western blotting.Results:The transfection rate of 35% was attained when FITC labeled negative siRNA was transfected into cells with siPORT~TM NeoFX~TM.Conclusion:The transfection activity of siRNA is observed,which provides a basis for finding the better method to check the rate of the transfection.
Cite
Citations (0)
To transfect nm23-H1 into the BcaCD885 cell lines in order to get safe high-efficiency and low-toxicity, and to find out whether nm23-H1 could affect the invasion and metastases ability of BcaCD885 cell lines.Lipofect was used to transfect nm23-H1 into BcaCD885 cell lines; immunohistochemistry was used to detect the difference expression of nm23-H1 between transfected and non-transfected cell lines; then transwell-room and wash way were used to detect the difference of invasion and metastases ability between transfected and non-transfected cell lines.PCMV-NEO-BAM system gave the stability expression of nm23-H1; there was significant different NDPKA expression between transfected and non-transfected BcaCD885 cell lines; the invasion and metastases ability of transfected BcaCD885 cell lines decreased obviously.nm23-H1 can inhibit the metastases of BcaCD885 cell lines significantly.
Cite
Citations (2)
Primary cell
Cationic polymerization
Cite
Citations (2)
目的研究极低频电磁场暴露对小鼠脑和肝组织c-fos mRNA水平的影响.方法将小鼠暴露于50 Hz、0.2 mT及50 Hz、6.0 mT的电磁场中,持续2周或4周;采用竞争性RT-PCR方法检测小鼠脑和肝脏c-fos mRNA水平.结果50 Hz、0.2 mT及50 Hz、6.0 mT电磁场暴露2周后,小鼠脑组织c-fos mRNA的水平上升为(0.017 8±0.007 6)amol/120 ng cDNA和(0.009 2±0.004 2)amol/120 ng cDNA,与对照组[(0.001 2±0.000 5)amol/120 ng cDNA]比较,差异有显著性(P＜0.05);肝组织c-fos mRNA的水平上升为(0.0117±0.005 5)amol/120ng cDNA和(0.014 8±0.016 2)amol/120 ng cDNA,与对照组[(0.000 5±0.000 5)amol/120 ng cDNA]比较,差异有显著性(P＜0.05).0.2 mT和6.0 mT电磁场暴露4周后,小鼠脑组织c-fos mRNA的水平上升为(0.010 0±0.005 4)amol/120 ng cDNA和(0.019 8±0.007 9)amol/120 ng cDNA,与对照组[(0.001 5±0.000 8)amol/120ng cDNA]比较,差异有显著性(P＜0.05);肝组织c-fos mRNA的水平分别上升为(0.017 3±0.012 2)amol/120ng cDNA和(0.013 3±0.009 0)amol/120 ng cDNA,而对照组无表达.结论50 Hz电磁场暴露引起小鼠脑和肝脏c-fos基因转录水平明显上调.
Cite
Citations (1)
We studied the relationship between ras transfection and the drug resistance phenotype in the MCF-10A cell line (a non-transformed immortalized human breast epithelial cell line).Parental (MCF-10A) and c-Ha-ras transfected (MCF-10A H) cell lines were tested for resistance to Doxorubicin, Maphosphamide and Cisplatinum, by evaluating the inhibition of [3H]-thymidine incorporation. Cells were also examined for the expression of different resistance mechanisms by immunocytochemical and RT-PCR methods.ras-transfected cells were much more resistant to Cis-platinum than parental ones. GST-pi the GST isoenzyme frequently involved in human cancers) increased only in transfected cells. No expression of any other resistance mechanisms (MDR, MRP, LRP) was found by immunocytochemistry either in parental or in transfected cells; nevertheless, the more sensitive RT-PCR tests detected mrp products in parental and in transfected cells: gene expression levels were low and equivalent in both cell lines.ras transfection can induce resistance to Cis-platinum by increasing GST-pi expression.
MCF-7
Cite
Citations (9)
We have studied different methods for efficient DNA transfection in neuroblastoma cell line. Here we report that the optimum transfection mode by nucleofection method yielded the maximum efficiency for SK-N-SH cells. To optimize transfection we selected different methods like electroporation, liposome mediated and PEI transfection. The DNA that was transfected was pmaxGFP plasmid (3.49kb). Our results suggest that the most efficient method for DNA transfection is very important to study gene expression and regulation by finding ways to deliver DNA into the difficult to transfect mammalian cells. The transfection efficiency by electroporation was ~60-70%, Lipofection was ~ 18-20% and PEI yielded <1%.
Nucleofection
Cite
Citations (0)