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[Effects of c-myc antisense oligodeoxynucleotide transfected by liposome on HL-60 cells]
1Fujian Institute of Hematology, Union Hospital of Fujian Medical University, Fuzhou 350001, China.
Objective:
To investigate a delivery system for administering antisense oligodeoxynucleotide (AS-ODN) to HL-60 cells and study the antitumor effects of c-myc antisense oligodeoxynucleotide.
Methods:
Human leukemia cell line HL-60 was transfected with a synthetic oligodeoxynucleotide(ODN) complementary to the 5' end region of c-myc gene. In addition, the changes of function and morphology were observed after transfection of cationic liposome.
Results:
HL-60 cells incubated with c-myc AS-ODN/lipofectin complex were about 50% decreased in the 4th day as compared with control cells (P < 0.05). About 56% of HL-60 cells exposed to c-myc AS-ODN/lipofectin complex were NBT-positive and showed morphological changes with differentiated phenotype compared with 29% of untreated cells. The expression of the c-myc in the treated cells decreased markedly. The levels of c-myc protein after treatment decreased from (80.00 +/- 3.02)% to (41.67 +/- 4.56)% (P < 0.01). c-myc AS-ODN induced HL-60 cells apoptosis which morphological changes of apoptotic cells were investigated by fluorescence microsopy. A ladder-like pattern of DNA fragmentation was demonstrated on agorose gel electrophoresis and 17% apoptotic cells were found with flow cytometry.
Conclusions:
Antisense oligodeoxynucleotide can suppress the malignant phenotype of HL-60 cells and enhancement of the effect was observed by adding lipofectin.
Insights
Antisense oligodeoxynucleotides (AS-ODN) effectively suppressed HL-60 leukemia cell growth and induced differentiation. Lipofectin enhanced AS-ODN delivery and antitumor effects, offering a promising cancer therapy approach.
Area of Science:
- Molecular Biology
- Cancer Research
- Drug Delivery Systems
Context:
- Human leukemia cell line HL-60 is a model for studying cancer.
- Antisense oligodeoxynucleotides (AS-ODN) target specific gene expression.
- Lipofectin is a cationic liposome used for gene delivery.
Purpose:
- To evaluate a delivery system for AS-ODN in HL-60 cells.
- To investigate the antitumor effects of c-myc AS-ODN.
- To assess the role of lipofectin in enhancing AS-ODN efficacy.
Summary:
- HL-60 cells treated with c-myc AS-ODN/lipofectin showed a significant decrease in proliferation (approx. 50%).
- Transfected cells exhibited differentiation markers (NBT positivity) and suppressed c-myc expression and protein levels.
- c-myc AS-ODN induced apoptosis in HL-60 cells, evidenced by DNA fragmentation and flow cytometry.
Impact:
- AS-ODN can suppress the malignant phenotype of leukemia cells.
- Lipofectin enhances the therapeutic efficacy of AS-ODN.
- This study provides a basis for AS-ODN-based cancer therapies.