半乳糖基修饰的反义硫代寡核苷酸对肝癌多药耐药细胞株MDR_1基因过度表达的抑制作用  

Inhibition of MDR1 Gene Overexpression of Human Hepatoma Resistant Cells by Galactose Modified Antisense Oligodeoxynucleotides

作　　者：郭军[1,2] 王宝成[1,2] 董冰 师秋丽[1,2] 狄剑时[1,2] 

机构地区：[1]济南军区总医院肿瘤治疗研究中心 [2]济南市第四人民医院免疫科

出　　处：《中国肿瘤临床》1998年第7期480-483,共4页Chinese Journal of Clinical Oncology

摘　　要：设计合成三种互补ＭＤＲ１ｍＲＮＡ的反义硫代寡核苷酸（ＡＳＯＤＮ），分别互补ＭＤＲ１ｍＲＮＡ序列起始区、含ＡＵＧ起始密码子区及针对Ｌｏｏｐ形成位点区的序列，作用于阿霉素（ＤＯＸ）诱导的肝癌多药耐药细胞株ＢＥＬ－７４０２／ＤＯＸ。经流式细胞分析及ＲＴ－ＰＣＲ的方法检测，发现三种ＡＳＯＤＮ均能不同程度地降低ＢＥＬ－７４０２／ＤＯＸ细胞膜表面Ｐ－ＧＰ含量；同时ＭＤＲ１ｍＲＮＡ的表达也有轻度降低，其中尤以互补ＭＤＲ１ｍＲ－ＮＡＬｏｏｐ形成位点的ＡＳＯＤＮ抑制作用最强。以导向配体Ｇａｌ１０－ＰＬＬ修饰此ＡＳＯＤＮ后，作用于ＢＥＬ－７４０２／ＤＯＸ细胞，发现与相同剂量未修饰ＡＳＯＤＮ比较，Ｐ－ＧＰ含量由７６．８％降至１９．８％；对ＡＤＭ的ＩＣ５０由１．２８μｇ／ｍｌ降至０．４２μｇ／ｍｌ。实验说明，半乳糖基修饰的互补ＭＤＲ１ｍＲＮＡＬｏｏｐ形成位点的ＡＳＯＤＮ能够明显降低肝癌多药耐药细胞膜表面Ｐ－ＧＰ的含量，并较大程度地逆转多药耐药细胞ＢＥＬ－７４０２／ＤＯＸ对化疗药物的耐受性。In order to reverse the MDR1 gene product Pglycoprotein mediated drug resistance in hepatoma resistant cells in a specific manner, three ASODNs towards different MDR1mRNA regions were synthesized and incubated with doxorubicin resistant hepatoma cells BEL7402/DOX. After 24h, 48h, 72h, cellular PGP levels were determined by immunoflowcytometry, and MDR1mRNA expression was evaluated using RTPCR after 72h. Then the sequence II ASODN was modified by galactose and the PGP level was determined with the same method. The sensitivity to ADM was assayed with MTT colorimetric technic. It was found that ADMs lowered the PGP level to varying degrees, and that it only slightly lowered the expression of MDR1mRNA. The modified ASODN was found even more efficient in lowering the PGP level and in reversing the BEL7402/DOX cell's drug resistance to ADM.

关 键 词：肝肿瘤 药物疗法 半乳糖 ASODN 多药耐药 

分 类 号：R735.705[医药卫生—肿瘤]