Related Experiment Videos
Human promyelocytic leukemia HL-60 cell proliferation and c-myc protein expression are inhibited by an antisense
E L Wickstrom1, T A Bacon, A Gonzalez
1Department of Chemistry, University of South Florida, Tampa 33620.
Abstract:
The human promyelocytic leukemia cell line HL-60 overexpresses the c-myc protooncogene. A calculated secondary structure for c-myc mRNA placed the initiation codon in a bulge of a weakly base-paired region. Treatment of HL-60 cells with 5' d(AACGTTGAGGGGCAT) 3', complementary to the initiation codon and the next four codons of c-myc mRNA, inhibited c-myc protein expression in a dose-dependent manner. However, treatment of HL-60 cells with 5' d(TTGGGATAACACTTA) 3', complementary to nucleotides 17-31 of vesicular stomatitis virus matrix protein mRNA, displayed no such effects. These results agree with analogous studies of normal human T lymphocytes [Heikkila, R., Schwab, G., Wickstrom, E., Loke, S. L., Pluznik, D. H., Watt, R. & Neckers, L. M. (1987) Nature (London) 328, 445-449], except that only one-third as much oligomer was needed for a comparable effect. Proliferation of HL-60 cells in culture was inhibited in a sequence-specific, dose-dependent manner by the c-myc-complementary oligomer, but neither the oligomer complementary to vesicular stomatitis virus matrix protein mRNA nor 5' d(CATTTCTTGCTCTCC) 3', complementary to nucleotides 5399-5413 of human immunodeficiency virus tat gene mRNA, inhibited proliferation. It thus appears that antisense oligodeoxynucleotides added to myc-transformed cells via culture medium are capable of eliciting sequence-specific, dose-dependent inhibition of c-myc protein expression and cell proliferation.
Insights
Antisense oligodeoxynucleotides targeting the c-myc gene inhibited its protein expression and cell proliferation in HL-60 leukemia cells. This sequence-specific inhibition was dose-dependent and more effective than in previous studies.
Area of Science:
- Molecular Biology
- Oncology
- Antisense Technology
Background:
- The HL-60 cell line overexpresses the c-myc protooncogene, a key factor in cell proliferation.
- The secondary structure of c-myc mRNA positions the initiation codon in a vulnerable region.
Purpose of the Study:
- To investigate the efficacy of antisense oligodeoxynucleotides in inhibiting c-myc protein expression and cell proliferation in HL-60 cells.
- To determine the sequence specificity and dose-dependency of this inhibition.
Main Methods:
- Treatment of HL-60 cells with specific antisense oligodeoxynucleotides complementary to c-myc mRNA.
- Treatment with control oligodeoxynucleotides targeting other mRNAs (vesicular stomatitis virus matrix protein, HIV tat gene).
- Assessment of c-myc protein expression and cell proliferation inhibition.
Main Results:
- A c-myc-specific antisense oligodeoxynucleotide significantly inhibited c-myc protein expression in a dose-dependent manner.
- This c-myc-targeting oligomer also inhibited HL-60 cell proliferation in a sequence-specific and dose-dependent manner.
- Control oligodeoxynucleotides showed no significant inhibitory effects on c-myc expression or cell proliferation.
Conclusions:
- Antisense oligodeoxynucleotides can effectively inhibit c-myc protein expression and proliferation in HL-60 cells.
- The observed inhibition is sequence-specific and dose-dependent.
- This approach demonstrates potential for therapeutic applications in c-myc-driven malignancies.