RPL6/Taxreb107在胃癌耐药细胞系SGC7901/ADR中的差异表达及其与MDR相关性的初步研究  

Differential expression of RPL6/Taxreb107 in drug resistant gastric cancer cell line SGC7901/ADR and its correlation with multiple-drug resistance

作　　者：杜静平[1] 金晓航[1] 时永全[1] 曹云新[2] 赵艳秋[1] 刘长江[1] 尹芳[1] 胡文华[1] 陈宝军[1] 乔泰东[1] 樊代明[1] 

机构地区：[1]第四军医大学西京医院消化病研究所,西安710032 [2]第四军医大学分子免疫学教研室

出　　处：《中华肿瘤杂志》2003年第1期21-25,共5页Chinese Journal of Oncology

基　　金：国家自然科学基金资助项目(30030140);创新研究群体科学基金资助项目(30024002)

摘　　要：目的探讨RPL6/Taxreb107在胃癌耐药细胞系SGC7901/ADR与胃癌细胞系SGC7901的差异表达,以及与胃癌多药耐药(MDR)的相关性。方法 提取SGC7901和SGC7901/ADR细胞系总RNA;采用内对照RT-PCR检测RPL6基因和Northern杂交、基因克隆与表达、真核表达载体的构建;以电穿孔法基因转染;以流式细胞仪检测瞬时转染细胞的阿霉素蓄积和潴留。结果 内对照RT-PCR和Northern杂交证实RPL6/Taxreb107在SGC7901/ADR中高表达。将PCR产物克隆入pUCm-T载体并经测序证实。构建正义和反义真核表达载体(pcDNA3.1)并经酶切鉴定证实后,以电穿孔法将正义真核表达载体转入SGC7901,反义真核表达载体转入SGC7901/ADR。转染48 h后检测转染细胞的阿霉素蓄积和潴留,结果提示RPL6/Taxreb107转入细胞后对细胞的耐药性有影响。结论 RPL6/Taxreb107在耐药胃癌细胞中高表达,对胃癌的MDR有影响。Objective To investigate the differential expression of RPL6/Taxrebl07 between drug-resistant gastric cancer cell line SGC7901/ADR and gastric cancer cell line SGC7901 as well as its correlation with multiple-drug resistance(MDR) in gastric cancer cells. Methods Total RNA was extracted from SGC7901 and SGC77901/ADR, with internal control RT-PCR, Northern blot, gene cloning and expression, construction of eukaryotic expression vector, gene transfection by electroporation. The accumulation and retention of ADR in transiently transfected cell was detected by flow cytometry. Results The internal control RT-PCR and Northern blot showed high RPL6/Taxrebl07 expression in SGC7901/ADR cell line. Sense and antisense eukaryonic expression vectors demonstrated by double enzyme digestion were successfully transfected into gastric cancer cell line SGC7901 and SGC7901/ADR respectively by electroporation. The accumulation and retention of ADR detected 48 hours after transfection showed that RPL6 gene had shown effect on drug resistance in gastric cancer cell. Conclusion The high expression of RPL6/Taxrebl07 in drug resistant gastric cancer cell shows its correlation with multiple-drug resistance in gastric cancer.

关 键 词：胃癌 多药耐药性 核糖体蛋白L6 SGC7901细胞 

分 类 号：R735.2[医药卫生—肿瘤]