Jian-Wei Jiang1, Yuan Zhang. 1. Institute of Hematology, Medical College, Jinan University, Guangzhou, Guangdong 510632, P.R. China.
Abstract
BACKGROUND & OBJECTIVE: Deliveries of c-myc antisense oligonucleotide (ASODN) mediated by liposome or adenvirus could suppress proliferation of human hepatocellular carcinoma cells, and tumor growth of mice hepatoma model. However, these deliveries lack targeting effects. Receptor-mediated drug delivery is being used in gene and ASODN delivery due to its high targeting efficiency. This study was to evaluate targeting delivery effect of c-myc ASODN mediated by galactose-polyethyleneimine (Gal-PEI) on human hepatocellular carcinoma cell line Bel-7402. METHODS: Bel-7402, and lymphoma cell lines, U937 and Raji, were cultured with fluorescence-labeled Gal-PEI-c-myc ASODN, and c-myc ASODN. The uptaking rates of Gal-PEI-c-myc ASODN, and intracellular mean fluorescence intensities of Bel-7402 and U937 cells were tested by flow cytometry. Morphology of Gal-PEI-c-myc ASODN and c-myc ASODN entering Bel-7402, U937, and Raji cells was observed under phase-contrast fluorescence microscope. Effects of Gal-PEI-c-myc ASODN of various concentrations on proliferation of these cells were detected by trypan blue dye method. RESULTS: After cultured for 10 min to 4 h, the uptaking rate, and intracellular mean fluorescence intensity of Gal-PEI-c-myc ASODN cultured Bel-7402 cells (88.25%-98.66%, and 38.61%-111.9%) were higher than those of c-myc ASODN cultured Bel-7402 cells, and Gal-PEI-c-myc ASODN cultured U937 cells, significant differences were detected by Poisson test (P < 0.01). Observed by phase-contrast fluorescence microscope, Gal-PEI-c-myc ASODN entered Bel-7402 cells effectively, but can't enter U937, and Raji cells effectively at the same concentration. C-myc ASODN can't enter Bel-7402 cells effectively at the same ASODN concentration. After incubation with Gal-PEI-c-myc ASODN (containing 0.25-1.25 micromol/L of c-myc ASODN) for 48 h, proliferation of Bel-7402 cells was significantly inhibited (P < 0.01=, but no significant differences were detected in U937, and Raji cells (P >0.05). CONCLUSION: Gal-PEI-c-myc ASODN has high targeting delivery effect on Bel-7402 cells, which enhances the intercellular concentration of c-myc ASODN effectively, but it has no such effects on U937, and Raji cells.
BACKGROUND & OBJECTIVE: Deliveries of c-myc antisense oligonucleotide (ASODN) mediated by liposome or adenvirus could suppress proliferation of humanhepatocellular carcinoma cells, and tumor growth of micehepatoma model. However, these deliveries lack targeting effects. Receptor-mediated drug delivery is being used in gene and ASODN delivery due to its high targeting efficiency. This study was to evaluate targeting delivery effect of c-mycASODN mediated by galactose-polyethyleneimine (Gal-PEI) on humanhepatocellular carcinoma cell line Bel-7402. METHODS:Bel-7402, and lymphoma cell lines, U937 and Raji, were cultured with fluorescence-labeled Gal-PEI-c-mycASODN, and c-mycASODN. The uptaking rates of Gal-PEI-c-mycASODN, and intracellular mean fluorescence intensities of Bel-7402 and U937 cells were tested by flow cytometry. Morphology of Gal-PEI-c-mycASODN and c-mycASODN entering Bel-7402, U937, and Raji cells was observed under phase-contrast fluorescence microscope. Effects of Gal-PEI-c-mycASODN of various concentrations on proliferation of these cells were detected by trypan blue dye method. RESULTS: After cultured for 10 min to 4 h, the uptaking rate, and intracellular mean fluorescence intensity of Gal-PEI-c-mycASODN cultured Bel-7402 cells (88.25%-98.66%, and 38.61%-111.9%) were higher than those of c-mycASODN cultured Bel-7402 cells, and Gal-PEI-c-mycASODN cultured U937 cells, significant differences were detected by Poisson test (P < 0.01). Observed by phase-contrast fluorescence microscope, Gal-PEI-c-mycASODN entered Bel-7402 cells effectively, but can't enter U937, and Raji cells effectively at the same concentration. C-mycASODN can't enter Bel-7402 cells effectively at the same ASODN concentration. After incubation with Gal-PEI-c-mycASODN (containing 0.25-1.25 micromol/L of c-mycASODN) for 48 h, proliferation of Bel-7402 cells was significantly inhibited (P < 0.01=, but no significant differences were detected in U937, and Raji cells (P >0.05). CONCLUSION:Gal-PEI-c-mycASODN has high targeting delivery effect on Bel-7402 cells, which enhances the intercellular concentration of c-mycASODN effectively, but it has no such effects on U937, and Raji cells.