Further Use of Protein Kinase N Beta
a technology of protein kinase and beta, which is applied in the field of use of protein kinase n beta, can solve the problems of a large threat to human health, numerous tumours and cancers, and still time and money-consuming
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Benefits of technology
Problems solved by technology
Method used
Image
Examples
example 1
Materials and Methods
[0147]Cell Culture
[0148]Human prostate carcinoma PC-3 cells were obtained from the American Type Culture Collection (ATCC). Cells were cultured in F12K Nutrient Mixture (Kaighn's modification) containing, 10% fetal calf serum (CS), gentamycin (50 μg / ml) and amphotericin (50 ng / ml).
[0149]Transfections were carried out in 96 well or 10-cm plates (at 30% to 50% confluency) by using various cationic lipids such as Oligofectamine, Lipofectamine (Life Technologies), Argfectin50 or Profectin50 (Atugen / GOT Berlin, Germany), or FuGene 6 (Roche) according to the manufacturer's instructions. GeneBlocs were transfected by adding pre-formed 5× concentrated complex of GeneBloc and lipid in serum-free medium to cells in complete medium. The total transfection volume was 100 μl for cells plated in 96 wells and 10 ml for cells in 10 cm plates. The final lipid concentration was 0.8 to 1.2 μg / ml depending on cell density; the GeneBloc concentration is indicated in each experiment....
example 2
Experimental Proof-of-Concept on the Suitability of Downstream Drug Targets
[0169]As outlined in the introductory part of this specification which is incorporated herein by reference, targets linked downstream to a signalling pathway are valuable for the design or development of both medicaments and diagnostic agents. It is obvious that, if the particular target is linked to different other pathways or due to its position within the signalling pathway is linked to a number of biological phenomena such as, e.g. metastasis and migration, growth translation apoptosis, cell cycle, DNA repair and the like as in the case of PI-3 kinase, any compound addressing this target is likely to have a number of side effects which may be detrimental to the system and undesired from the medical point of view. Accordingly, targets that act further downstream should be the first choice for therapeutic intervention.
[0170]The present inventors have found that under the control of the PI 3-kinase pathway f...
example 3
Identification of PKN Beta as Downstream Drug Target within the PI 3-Kinase Pathway
[0176]The basic experimental approach is shown in FIG. 3. PC3 cells grown on Matrigel were either treated with DMSO or the PI 3-K inhibitor LY294002 and total RNA was isolated from each sample. Differential Affymetrix gene expression profiling was performed and expression was confirmed using real time RT-PCR Taqman assay. p110α was used as a non-differential standard. PC3 cells are PTEN− / − which means that the tumor suppressor PTEN is factually lacking in these cells so that the PI 3-kinase pathway is permanently activated which leads to an increased metastatic activity or behaviour of the cells which is expressed by their growth pattern in the matrigel assay. Cells with invasive growth potential exhibit enhanced growth on basement membrane such as matrigel matrix. (Petersen, O. W., Ronnov-Jessen, L., Howlett, A. R. and Bissell, M. J. (1992) Interaction with basement membrane serves to rapidly disting...
PUM
| Property | Measurement | Unit |
|---|---|---|
| molecular weight | aaaaa | aaaaa |
| transfection volume | aaaaa | aaaaa |
| transfection volume | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
Login to View More