Inhibition of Multidrug Resistance in Human Tumor Cell by Naphthyl Imide-Conjugated Antisense Oligonucleotide
10.3321/j.issn:1000-467X.2001.04.007
- VernacularTitle:反义寡核苷酸-萘二酰亚胺偶联物对多药耐药性肿瘤细胞的抑制作用
- Author:
Jun-Sheng LI
1
;
Dong-Zhi WEI
;
Yu-Xun ZHOU
;
Jian-Chao QIAN
;
Dong-Hui ZHU
;
Yuan-Xing ZHANG
Author Information
1. 华东理工大学
- From:Chinese Journal of Cancer
2001;20(4):373-378
- CountryChina
- Language:Chinese
-
Abstract:
Objective： This study was designed to investigate the effectiveness of natural and naphthyl imide-conjugated antisense oligonucleotide on inhibiting mdr1 gene expression and of overcome the problem of multidrug resistance in human epidemic carcinomata anti-adriamycin cells(KB-A-1). Methods： A novel naphthyl imide-conjugated mdr1 antisense oligonucleotide was synthesized by S-alkylation reaction. Compared with the natural one, the effectiveness of naphthyl imide-conjugated mdr1 antisense oligonucleotide on inhibiting the multidrug resistance was detected by MTT colometric assay and ELISA. Results： The abilities of antisense oligonucleotides to inhibit the cell growth of KB-A-1 mainly depended upon their targeted sequences selected. The inhibiting rates to KB-A-1 cells of antisense oligonucleotides (from Oligo Ⅰ to Oligo Ⅳ ) which bind the targeted sequences of the translation initiation site of mdr1 were 13.34％ ,14.32％ ,26 00％ ， and 25.37％ ， respectively. Among four oligonucleotides,the activity of Oligo Ⅲ to inhibit the growth of KB-A-1 cell was the highest (P<0.01). Compared with the unmodified one, the inhibition of antisense oligonucleotides to the growth of KB-A-1 cell was increased by conjugating with naphthyl imide. The ability of naphthyl imide-conjugated antisense oligonucleotide to resist serum-mediated nuclease was also increased as demonstrated by gel shift electrophoresis. The antisense oligonucleotide or its naphythyl imide conjugate could only inhibit the growth of KB-A-1 cells and had no effect on the growth of KB-3-1 cells. So it could be inferred that the inhibition of KB-A-1 cells was due to the repression of gene expression of mdr1 in KB-A-1 cells by antisense oligonucleotide or its naphthyl imide conjugate. ELISA showed that the P-glycoprotein expressions were more strongly inhibited by naphthyl imide-conjugated oligonucleotide than that by unmodified oligonucleotide. Conclusion: Inhibition of human tumor cell growth was due to the inhibition on mdr1 gene expression and the reversal of multidrug resistance by antisense oligonucleotide and its naphthyl imide conjugated. 1, 8-Naphthyl imide could improve the properties of natural oligonucleotide in inhibiting the multdrug resistance and resisting nuclease by covalently linkage at the end of oligonucleotide.
