Bound antibody was detected using enhanced chemiluminescence's reagent (ECL Western Blotting System; Amersham Bioscience Corp., Piscataway, NJ, USA).
To determine if apoptotic cells are observed in treated cells, we performed the terminal deoxynucleotidyl transferase-mediated d UTP nick end label (TUNEL) assay using Apop Tag kit (Invitrogen) as described previously (Komata et al, 2000).
A control antibody, Ig G from goat serum was purchased from SIGMA (St Louise, MO, USA).
Therefore, using the GFP-LC3 expression vector kindly supplied by Dr N Mizushima and Dr T Yoshimori, (National Institute for Basic Biology, Japan), U87-MG cells expressing GFP-LC3 (GFP-LC3-U87-MG cells) were established.Activated recombinant Akt was purchased from Upstate Biotechnology (Lake Placid, NY, USA). anzeigen sie sucht ihn Hamm Human malignant glioma U87-MG and T98G cells and human normal fibroblasts MRC5 were purchased from American Tissue Culture Collection (Rockville, MD, USA).In the present study, using neutralising antibody to PDGF, we investigated the effect of the inhibition of PDGF signalling on malignant glioma cells with high levels of PDGF and fibroblast cells with low levels of PDGF.We demonstrate that the inhibition of PDGF signalling induces autophagy, but not apoptosis in tumour cells, while fibroblast cells are resistant to treatment with anti-PDGF neutralising antibody.
Dating gifte Langeland
Our results suggest a novel function of disruption of PDGF signalling on tumour cells.Anti-human PDGF neutralising antibody (goat) was purchased from R&D systems (Minneapolis, MN, USA).Obviously, a novel treatment strategy of this tumour is required for prolonged survival.Malignant tumour cells are well characterised by the unfettered reproduction through cell division of their progeny and themselves.Department of Neurosurgery, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Blvd., Unit 64, Houston, TX 77030, USACorrespondence: Dr S Kondo, E-mail: [email protected] 31 July 2003; Revised 21 November 2003; Accepted 28 November 2003receptor complex represents a promising strategy for the treatment of malignant gliomas.
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