Effects of antisense-myc-conjugated single-walled carbon nanotubes on HL-60 cells.
Antisense therapy is limited in application in clinical therapy because of two existing problems: rapid degradation of antisense nucleic acids and poor diffusion across the cell membrane. Here we report the use of single-walled carbon nanotubes as delivery system for transporting antisense myc into HL-60 cells. Antisense-myc-conjugated single-walled carbon nanotubes were synthesized, characterized by atomic force microscopy (AFM), fluorescent microscopy, and Raman spectroscopy. Incubated with HL-60 cells at 37 degrees C, the single-walled carbon nanotubes (SWNTs) inside HL-60 cells were measured quantitatively by HPLC. Expression of c-myc gene and protein were analyzed by reverse transcriptase PCR and Western blot, respectively. Results showed that as-myc-conjugated SWNTs dropped character peaks at quadruple wavenumber (cm(-1)) compared with SWNTs, and could enter into HL-60 cells within 15 min after incubation, whose uptake amounts by HL-60 cells increased as the incubation time increased. After 48 hours, the amount of SWNTs in HL-60 cells began to decrease. Compared with as-myc and SWNTs, as-myc-conjuagted SWNTs exhibited the strongest inhibition on the proliferation of HL-60 cells, induced cell apoptosis, and down-regulated lowest expression of c-myc gene and C-MYC protein. The SWNTs can directly deliver as-myc into HL-60 cells, enhance the inhibition of as-myc on HL-60 cells, and is likely a better delivery system for antisense therapy. Antisense modified SWNTs can be used for intracellular gene regulation with potential applications in tumor therapy and drug delivery.
Cui
Daxiang
D
Bio-X DNA Computer Consortium, Key Laboratory for Thin Film and Microfabrication of Ministry of Education, Institute of Micro and Nano Science Technology, Shanghai JiaoTong University, 1954 Huashan Road, Shanghai 200030, P. R. China.
Tian
Furong
F
Coyer
Sean R
SR
Wang
Jicun
J
Pan
Bifeng
B
Gao
Feng
F
He
Rong
R
Zhang
Yafei
Y
D045744:Cell Line, Tumor; D002851:Chromatography, High Pressure Liquid; D018922:HL-60 Cells; D006801:Humans; D018625:Microscopy, Atomic Force; D036103:Nanotechnology / Q000379:methods*; D037742:Nanotubes, Carbon / Q000737:chemistry*; D016376:Oligonucleotides, Antisense / Q000737:chemistry*; D016133:Polymerase Chain Reaction; D016271:Proto-Oncogene Proteins c-myc / Q000737:chemistry*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013696:Temperature; D013997:Time Factors
D037742:Nanotubes, Carbon; D016376:Oligonucleotides, Antisense; D016271:Proto-Oncogene Proteins c-myc