反义肽核酸逆转人神经母细胞瘤多药耐药性的实验研究  

Reversion of multidrug resistance of human neuroblastoma by antisense peptide nucleic acid

作　　者：郭华[1] 张庆林[1] 王成伟[1] 张健[1] 孔建新[1] 刘福生[1] 马道新[1] 卞继峰[1] 

机构地区：[1]山东大学第二医院神经外科,济南山东省立医院神经外科250033

出　　处：《中华神经外科杂志》2005年第2期113-117,共5页Chinese Journal of Neurosurgery

基　　金：国家自然科学基金资助项目(30070272)

摘　　要：　目的　探讨反义肽核酸(PNA)的细胞转染方法及对体外培养的神经母细胞瘤多药耐药性(MDR)的逆转作用。方法　以人MDR 1基因mRNA为靶点设计合成两反义PNA序列,利用PNA DNA杂交,阳离子脂质体介导转染神经母细胞瘤耐药细胞株SK N SH。应用流式细胞术、RT-PCR、MTT等方法分别检测反义PNA的转染效率、转染前后细胞P 糖蛋白(P gp)、MDR 1基因mRNA的表达以及对化疗药物的敏感性。结果　荧光标记的反义PNA转染肿瘤细胞后,细胞平均荧光强度显著增强,且呈浓度依赖性。两反义PNA均使SK N SH细胞P gp表达明显降低,阿霉素和长春新碱对细胞的半数抑制浓度值明显下降,MDR 1mRNA表达轻度降低。而PNA1转染对照组C6 /adr细胞后,上述变化不明显。结论　PNA DNA杂交阳离子脂质体转染法可有效增加细胞对PNA的摄取,特异序列的反义PNA可有效逆转神经母细胞瘤的MDR特性。Objective To investigate the efficiency of a peptide nucleic acid (PNA) delivery system by using liposome via PNA-DNA hybrids. And to test the effectiveness of antisense PNA in reversing multidrug resistance (MDR) in human neuroblastoma cell line SK-N-SH. Methods Two antisense PNAs were designed targeting at the MDR-1 mRNA AUG initiation region and 5′-UTR region. Two PNAs were combined with partially complement DNAs respectively and then delivered into cells using cationic liposome. The transfection efficiency, change of sensitivity to chemotherapy drugs, expression of P-glycoprotein (P-gp) and MDR-1 mRNA were measured by flow cytometry, MTT and RT-PCR respectively. Results Transfection of PNA increased the cell average fluorescence intensity significantly and the extent of increase was dependent on the concentration of PNA. After transfection by both PNAs, P-gp expression and IC50 to ADM and VCR of SK-N-SH cells decreased significantly. But for C6/adr cells as control, PNA1 did not cause such changes. There was slight decrease of the level of MDR-1 mRNA expression after transfection of SK-N-SH cells by both PNAs, but not significant. Conclusions PNA can be delivered into tumor cells in form of PNA-DNA hybrids by cationic liposome. Properly designed antisense PNA can reverse the MDR of SK-N-SH cells efficiently and specifically.

关 键 词：神经母细胞瘤 反义肽 转染 逆转 多药耐药性 P-GP MDR 结论 方法 目的 

分 类 号：R73-3[医药卫生—肿瘤]