Jiang Jian-Wei, Zhang Yuan
Institute of Hematology, Medical College, Jinan University, Guangzhou, Guangdong 510632, P.R. China.
Ai Zheng. 2004 Nov;23(11):1288-93.
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.
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.
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).
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.
脂质体或腺病毒介导的c-myc反义寡核苷酸（ASODN）可抑制人肝癌细胞增殖及小鼠肝癌模型肿瘤生长。然而，这些递送方式缺乏靶向作用。受体介导的药物递送因其高靶向效率正被用于基因和ASODN递送。本研究旨在评估半乳糖-聚乙烯亚胺（Gal-PEI）介导的c-myc ASODN对人肝癌细胞系Bel-7402的靶向递送效果。
用荧光标记的Gal-PEI-c-myc ASODN及c-myc ASODN培养Bel-7402、淋巴瘤细胞系U937和Raji。采用流式细胞术检测Gal-PEI-c-myc ASODN的摄取率及Bel-7402和U937细胞的细胞内平均荧光强度。在相差荧光显微镜下观察Gal-PEI-c-myc ASODN及c-myc ASODN进入Bel-7402、U937和Raji细胞的形态。采用台盼蓝染色法检测不同浓度Gal-PEI-c-myc ASODN对这些细胞增殖的影响。
培养10分钟至4小时后，Gal-PEI-c-myc ASODN培养的Bel-7402细胞的摄取率及细胞内平均荧光强度（88.25%-98.66%，38.61%-111.9%）高于c-myc ASODN培养的Bel-7402细胞及Gal-PEI-c-myc ASODN培养的U937细胞，经泊松检验差异有统计学意义（P<0.01）。相差荧光显微镜观察显示，Gal-PEI-c-myc ASODN能有效进入Bel-7402细胞，但相同浓度下不能有效进入U937和Raji细胞。相同ASODN浓度下，c-myc ASODN不能有效进入Bel-7402细胞。Gal-PEI-c-myc ASODN（含0.25-1.25μmol/L的c-myc ASODN）孵育48小时后，Bel-7402细胞增殖受到显著抑制（P<0.01），但U937和Raji细胞无显著差异（P>0.05）。
Gal-PEI-c-myc ASODN对Bel-7402细胞具有高靶向递送效果，能有效提高c-myc ASODN在细胞内的浓度，但对U937和Raji细胞无此作用。
