Quantification intracellular levels of oligodeoxynucleotide-doxorubicin conjugate in human carcinoma cells in situ.
Bifunctional oligodeoxynucleotide-intercalator conjugates provide new tools for a selective control of gene expression and may have potential applications as novel therapeutic agents. An 'in situ' analysis method has been developed to determine intracellular levels of oligodeoxynucleotide- doxorubicin conjugate which may be used to reverse the multidrug resistance in KB-A-1 cells. A linear response over a broad concentration range (0.05-6.0 microM, r2 = 0.97) was obtained. The limit of detection and quantitation was set at 0.015 and 0.05 microM, respectively. Assay validation studies revealed that compared with HPLC, the method is rapid, high simple, high throughput and convenient for the determination of the conjugate in cells in situ. The uptake studies of the conjugate in cells using this method demonstrated that the cellular accumulation of the conjugate in KB-A-1 cells was dependent on a dynamic balance between influx and efflux processes.
Ren
Yuhong
Y
State Key Laboratory of Bioreactor Engineering, Institute of Biochemistry, East China University of Science and Technology, Shanghai 200237, China.
Wei
Dongzhi
D
D002277:Carcinoma / Q000737:chemistry*; D045744:Cell Line, Tumor; D002851:Chromatography, High Pressure Liquid; D004317:Doxorubicin / Q000032:analysis*; D006801:Humans; D009841:Oligonucleotides / Q000032:analysis*; D012015:Reference Standards; D015203:Reproducibility of Results; D012996:Solutions; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet
D009841:Oligonucleotides; D012996:Solutions; D004317:Doxorubicin