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MessagePosté le: Jeu 25 Sep - 07:18 (2014) Sujet du message: As expected, after a 48 h treat ment period, OcTMAB induced Répondre en citant

 Effect of cassini expression Our experiments showed that induction of cassini RNA in cells correlates with specific stress stimuli, but our data do not provide information on whether the increased levels are detrimental to the cells or contribute a survival advantage. Because of the abundance of this RNA upon stress of cells, and the possibility that other transcribed loci exist in the genome INNO-406 構造 with identical se quence, ablation of the RNA using siRNA does not ap pear technically feasible. We therefore used transient transfection in 293 cells to attempt to achieve expression levels that would significantly add to that already induced endogenously by stress, and also subjected the cells to a short term treatment with chemotherapeutic drugs.

The results of these experiments rule out the pos sibility that cassini induction correlates with or contri butes Lapatinib 溶解度 to cell death, and suggest that high levels of this RNA provide some form of protection against the cyto static activity of such drugs. Conclusions We conclude that satellite repeat sequences are widely conserved and, as shown here, their transcription can be specifically regulated. Our analysis of cassini demon strates that among this large family of loci, some are likely to participate in the response of acute lympho blastic leukemia cells to cytotoxic drug treatment in can cer therapy in the presence of stromal support, and that this may provide some protection against these drugs. Methods Data access The microarray data have been deposited in GEO under accession number GSE33329.

Cell culture and experimental design 8093 cells, a murine pro B ALL cell line that was isolated from a BCR/ABL transgenic mouse, human Ph negative US7 and Ph positive TXL2 cells LY2109761 TGF-beta/Smad 阻害剤 have been pre viously described in detail. Briefly, 8093 cells were cultured in McCoys 5A medium supple mented with 15% fetal bovine serum, 1% Gluta max, HEPES, 0. 1% beta mercaptoethanol, 1% penicillin/ streptomycin, and 0. 01 ng/mL of IL 3. US7 and TXL2R were cultured in MEM supplemented with 20% FBS, 1% Glutamax, and 1% penicillin/streptomycin. TXL2R cells were derived from TXL2 by prolonged cul ture in the presence of nilotinib. Cells are able to prolif erate normally in the presence of 500 nM of the drug. Primary total mouse splenocytes were incubated for 5 hrs in RPMI supplemented with 10% FBS, 1% Glutamax and 1 ug/mL of E. coli endotoxin or 1xPBS.

For drug treatment experiments, murine and human ALL cells were co cultured, directly or indirectly, in the presence of mitotically inactivated stromal MEF or OP9 feeder layers, respectively. For collection of leukemic cells in Figures 4 and 5A, a direct co culture was used that results in cell cell contact. Although ALL cells migrate underneath the stromal layer, they also pro liferate abundantly in the supernatant. Because the MEFs are fully adhered to the plates, a pure population of leukemic cells is available for isolation. In indirect co cuture, leukemic cells are cultured in Transwells with 0. 4 uM pore size, which pre cludes cell migration, but allows for free exchange of co culture media. The Transwells are placed over the MEF stromal layer, allowing for a clean isolation of the MEFs. Drugs including nilotinib, vincristine, and/or ASA, were refreshed every 48 hrs.


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MessagePosté le: Jeu 25 Sep - 07:18 (2014) Sujet du message: Publicité

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