J Korean Med Sci.  2003 Apr;18(2):184-190. 10.3346/jkms.2003.18.2.184.

Myeloablative Treatment Supported by Autologous Stem Cell Infusion with Neuroblastoma

• Ryu KH
• Seoh JY
• Jang PS
• Kim CW
• Koh SH
• Shin HY
• Ahn HS

• KMID: 2157620
• DOI: http://doi.org/10.3346/jkms.2003.18.2.184

Abstract

Bcr-abl antisense oligodeoxynucleotides (AS-ODNs) have provided evidence of an antileukemia effect when tested in vitro against Philadelphia-positive cells. In order to investigate the efficacy of AS-ODNs as purging agents in chronic myeloid leukemia (CML) patients, K562 cells, a human CML cell line, were treated in vitro with various types of AS-ODNs and interferon-alpha. Cells were treated in vitro for 0 and 36 hr with 40 microgram/mL of AS-ODNs, respectively, and incubated at 37 degrees C for 36 hr. Cytotoxic effects were measured by counting the number of viable cells as well as by MTT test. Clonogenic activities were evaluated by methylcellulose culture for 2 weeks. The effects of purging agents on the rearrangement of bcrabl gene were evaluated by RT-PCR. AS-ODNs inhibited the proliferation of K562 cells with time in cell count assay and MTT test. AS-ODNs were superior to INF-alpha in inhibiting clonogenic activity (recovery rate; 26.3% vs 64.0%). After incubation with bcr-abl AS-ODNs primers and mRNA isolated from K562 cells, positive bands were abolished, especially of b3a2 type and phosphorothioate type. Our results suggest that AS-ODNs mediated purging may be one of the efficient methods and that autograft may be an alternative treatment for allograft in high-risk group patients of CML if they do not have a stem cell donor.

Keyword

Oligonucleotides, Antisense; Leukemia, Myeloid, Chronic; Philadephia Chromosome

MeSH Terms

Bone Marrow Purging*
Colony-Forming Units Assay
Fusion Proteins, bcr-abl/genetics
Fusion Proteins, bcr-abl/metabolism
Hematopoietic Stem Cells/physiology*
Human
Leukemia, Myeloid, Chronic/therapy
Neuroblastoma/therapy*
Oligonucleotides, Antisense/metabolism*
Oligonucleotides, Antisense/therapeutic use
Transplantation, Autologous*
Tumor Cells, Cultured