Quantitation of resistance to cytosine arabinoside by myeloid leukemic cells expressing bcl-2

Eur J Haematol. 1996 Aug;57(2):149-56. doi: 10.1111/j.1600-0609.1996.tb01353.x.

Abstract

The presence of bcl-2 in myeloid leukemias has been associated with a decrease in therapy-induced apoptosis, reduced patient survival and in vitro autonomous growth of leukemic cells. The present study focuses on the quantitation of resistance to increasing doses of 1-beta-d-arabinofuranosylcytosine (Ara-C) by using hematological tumors expressing different levels of bcl-2. Scanning densitometry of Western blots demonstrated that the myeloid U-937 cells express low levels of bcl-2 (RD = 0.008), whereas the follicular lymphoma RL-7 expressed very high levels (RD = 3.084). Colony formation was also examined following incubation with Ara-C and RL-7 cells demonstrated a higher clonogenic survival (LD50 = 0.5 microns) when compared with U-937 cells (LD50 = 0.005 microM). Similarly, the level of bcl-2 expression in each cell line was also related to apoptosis with U-937 cells demonstrating increased DNA fragmentation when compared with RL-7 cells. To further evaluate the effect of upregulated bcl-2 on Ara-C treatment, U-937 cells were transfected with a retroviral vector carrying the murine bcl-2 or vector alone. Upregulation of bcl-2 by myeloid leukemic cells increased the resistance by 3 logs to Ara-C when comparing LD50 values from clonogenic assays, and decreased apoptosis by at least 3 logs when measuring dUTP positive cells by flow cytometry.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Antimetabolites, Antineoplastic / pharmacology*
  • Apoptosis / drug effects
  • Cytarabine / pharmacology*
  • DNA Fragmentation
  • DNA, Complementary / genetics
  • Drug Resistance, Neoplasm / genetics
  • Gene Expression Regulation, Neoplastic
  • Humans
  • Leukemia, Myeloid / genetics
  • Leukemia, Myeloid / pathology*
  • Lymphoma, Follicular / genetics
  • Lymphoma, Follicular / pathology*
  • Lymphoma, Large B-Cell, Diffuse / genetics
  • Lymphoma, Large B-Cell, Diffuse / pathology*
  • Mice
  • Neoplasm Proteins / analysis*
  • Neoplasm Proteins / biosynthesis
  • Neoplasm Proteins / genetics
  • Neoplastic Stem Cells / drug effects*
  • Proto-Oncogene Proteins c-bcl-2 / analysis*
  • Proto-Oncogene Proteins c-bcl-2 / biosynthesis
  • Proto-Oncogene Proteins c-bcl-2 / genetics
  • Transfection
  • Tumor Cells, Cultured / drug effects

Substances

  • Antimetabolites, Antineoplastic
  • DNA, Complementary
  • Neoplasm Proteins
  • Proto-Oncogene Proteins c-bcl-2
  • Cytarabine