Gene expression profiling of B cells from p80HT mice To gain a molecular unders

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 Gene expression profiling of B cells from p80HT mice To gain a molecular unders Empty Gene expression profiling of B cells from p80HT mice To gain a molecular unders

Post  huwan123456 on Tue Apr 08, 2014 7:28 am

DAPI was added to the final wash and the coverslips were mounted using Prolong Gold Antifade, Confocal images were ac quired using a Zeiss LSM 510 microscope. Analysis of tumor xenografts All animal procedures were performed in strict accordance with the NIH Guide for the Care ARN-509 臨床試験 and Use of Laboratory Animals and approved by the Institutional Animal Care and Use Committee at Dartmouth. To generate tumor xe nografts, 2 × 106 AsPC 1 or MiaPaCa 2 pancreas cancer cells were injected into the flanks of athymic nu nu mice. Drug treatments began after the tumors had reached 100 mm3. Gemcitabine was administered at 150 mg kg i. p. in phosphate buffered saline while MK 8776 was adminis tered at 50 mg kg i. p.

in B cyclodextrin, 45% w v solution in water, These doses were se lected based on a prior publication with these agents, The schedules of administration varied with experiment and are described in the results. Tumors were measured AUY922 臨床試験 with calipers in two dimensions and volume calculated based on the equation volume π 6 × length × width2. The comparisons between groups at each time point were made using a students t test for unpaired samples. The tests were two sided and a change with a p value 0. 05 was considered statistically significant. Some tumors were harvested, fixed in formalin, and serial sections were stained with anti Ki67 and anti geminin in the Research Pathology Shared Resource. For each tumor, at least 2 fields from each of 2 sections were photographed, each field represent ing about 1000 cells, 2 4 individual tumors were scored at each time point.

The number of cells positive for geminin was expressed as a percentage of those positive for Ki67. Results Impact of MK 8776 on gemcitabine induced cytotoxicity We previously analyzed MDA MB 231 and MCF10A cell lines for sensitivity ALK 阻害剤 to gemcitabine alone or when combined with MK 8776, This analysis has now been expanded to a large panel of cell lines, In this assay, cells were incubated with drugs for 24 h, and cell growth was then assessed after an additional 6 7 days. The results are expressed as the IC50 for gemcitabine alone or when incubated with low or high MK 8776, these concentrations were selected based on our prior experience showing differential sen sitivity of cell lines to this drug, The cells exhibit a wide range of sensitivity to gemcitabine alone, but concurrent incubation with 2 umol L MK 8776 resulted in an IC50 of 6.

5 nmol L for all the cell lines. This reflected a 4 66 fold sensitization to gemcitabine. We previously noted that some cell lines are particularly sensitive to MK 8776 alone, these in cluded U2OS, A498 and TK10, Our expanded screen has now identified AsPC 1 as sensitive to MK 8776, Most of the other cell lines tolerated 10 umol L MK 8776 for 24 h. For the sensitive cell lines, it was not possible to determine an IC50 for gemcitabine in combin ation with 2 umol L MK 8776. However in these cell lines sensitization was still observed when combined with 200 nmol L MK 8776. TK10 cells are an exception in this regard as they are very sensitive to gemcitabine alone so were not sensitized further.

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