摘要
目的 探讨反义策略 (反义核酸和Ribozyme)对肺腺癌细胞多药耐药的逆转效果。方法 分别以mdr1cDNA 9～ + 6和mdr1mRNA 880位点的GUC为靶位点 ,通过脂质体介导将相应的反义核酸 (ATCCATCCCGACCTC)和anti mdr1mdr1Ribozyme表达质粒DNA(pHβApr 1neo/mdr1 Rb )导入肺腺癌耐药细胞株A5 49/R ,分别采用RT PCR、流式细胞仪、罗丹明试验及MTT方法观察细胞的mdr1mRNA、Pgp表达、罗丹明的聚集和对阿霉素的敏感性的变化。结果 经反义核酸和anti mdr1mdr1Ribozyme处理的细胞mdr1mRNA、Pgp明显降低 ,细胞内罗丹明聚集 ,对阿霉素的敏感性分别提高 2 0倍和 180倍。结论 反义核酸和anti mdr1mdr1Ri bozyme具有强大的逆转肺癌MDR的作用 ,其作用水平在mRNA或DNA水平 ,使Pgp的表达受到强烈抑制 ,显示了良好的应用情景。
Objective To investigate the possibility of reversion of multidrug resistance in lung adenocarcinoma resistant cell line A549/R by antisense strategy including mdr1 antisense oligodeoxynucleotide and Ribozyme. Methods The target points, 9 to +6 in mdr1 cDNA sequence and GUC at 880 site of mdr1 mRNA were selected. Phosphorothioate antisense oligodeoxynucleotide and DNA of plasmid expressing anti mdr1 Ribozyme(pHβApr 1 neo/mdr1 Rb )corresponding to above target points were transfered into A549/R cell by lipofectin. The expression of mdr1 mRNA and Pgp, cellular rhodamine accumulation and sensitivity to doxorubicin were detected respectively in transfered cells and control cells by RT PCR, flow cytometry, rhodamine test and MTT. Results Treatment of A549/R cell with antisense oligodeoxynucleotide and Ribozyme led to decrease of mdr1 mRNA and Pgp expression, increase of rhodamine cellular accumulation, and 20 180 fold increase of sensitivity to doxorubicin compared to control cell. Conclusion The mdr1 antisense oligodeoxynucleotide and Ribozyme are possessed with powerful effect on reversion of MDR in lung adenocarcinoma resistant cell A549/R by inhibiting mdr1 transcription, cleaving mdr1 mRNA and decreasing Pgp expression. Antisense strategy is a promising method of reversion of multidrug resistance in lung cancer.
出处
《中国肺癌杂志》
CAS
2001年第3期165-168,共4页
Chinese Journal of Lung Cancer
关键词
肺腺癌
多药耐药性
反义核酸
RIBOZYME
Lung adenocarcinoma Multidrug resistance Antisense oligodeoxynucleotide Ribozyme
