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多药耐药MDRl基因的过度表达是导致肿瘤细胞产生MDR的重要机制.c-fos基因表达的变化很可能对MDRl基因的表达起着调控作用.为逆转肝癌多药耐药细胞对多种化疗药物的耐受性,本文分别设计合成了针对c-fos Exonl(914-938)的25聚反义寡核苷酸(ASODN),其序列为5’CAGCGGGAGGATCACGCCTCGTAGT-3’.同时设计合成了切割MDRl mRNA第196密码子GUC序列的锤头状Ribozyme,其双链序列为:5’-pGATCCATTAATACGCTCACTATAGAATCTTCGACTGATGAG-3’,和5’-pAATTCTCTTTCA GTTTCGTC-CTCACGGACTCATCAGAATG-3’.将Ribozyme基因定向克隆于逆转录病载体pDOR-neo的BamH Ⅰ位点,经病毒包装细胞PA317包装后感染人肝癌多药耐药细胞株BEL-7402/DOX,用c-fox ASODN 作用于此转化细胞株,48小时后流式细胞技术(FCM)检测联合治疗组细胞P-gp的表达为6.2～7.8%;单纯Ribozyme治疗组细胞为38～45%;对照组为93.4～97.5%.
Overexpression of multidrug resistance MDR1 gene is an important mechanism leading to MDR in tumor cells. The change of c-fos gene expression is likely to regulate the expression of MDR1 gene. To reverse the multidrug resistance of hepatocellular carcinoma cells against multiple chemotherapy Tolerability of the drug, this article designed and synthesized a 25-polyantisense oligonucleotide (ASODN) against c-fos Exonl (914-938), whose sequence is 5’CAGCGGGAGGATCACGCCTCGTAGT-3’. Simultaneously, the design cuts MDR1. Hammerhead Ribozyme of mRNA codon GUC sequence with double-stranded sequences: 5’-pGATCCATTAATACGCTCACTATAGAATCTTCGACTGATGAG-3’, and 5’-pAATTCTCTTTCAGTTTCGTC-CTCACGGACTCATCAGAATG-3’. Targeted Ribozyme gene cloned in retroviral vector pDOR -neo BamH I site, infected with viral packaging cells PA317, infected human hepatocellular carcinoma multidrug resistant cell line BEL-7402/DOX, and used c-fox ASODN to act on this transformed cell line, 48 hours later by flow cytometry ( The expression of P-gp in the combined treatment group was 6.2-7.8% in FCM and 38-45% in Ribozyme alone group and 93.4-97.5% in control group.