摘要
目的:对Ro 60变异体1在胃癌多药耐药中的功能进行研究.方法:克隆Ro 60变异体1编码基因,构建Ro 60变异体1编码基因的正义真核表达载体,将其转导入SGC7901细胞,应用半定量RT-PCR技术,对基因转染细胞进行鉴定,通过MTT法进行体外药物敏感性分析,借助流式细胞仪检测细胞内蓄积的阿霉素.结果:成功构建Ro 60变异体1正义真核表达载体:应用脂质体介导法将其转导入SGC7901,其表达量明显增加;体外药物敏感性试验提示,与SGC7901和SGC7901-pcDNA3.1细胞相比,转染细胞对长春新碱(IC_(50):2.87±0.10 mg/L vs 0.47±0.07 mg/L,0.63±0.08 mg/L,P<0.01)、5-氟尿嘧啶(IC_(50):3.89±0.12 mg/L vs 0.59±0.17 mg/L,0.92±0.12 mg/L,P<0.01)、丝裂霉素(IC_(50):1.02±0.06 mg/L vs 0.50±0.04 mg/L,0.73±0.09 mg/L,P<0.05)、顺铂(IC_(50):1.15±0.06 mg/L vs 0.46±0.04 mg/L.0.52±0.05 mg/L,P<0.01)、阿霉素(IC_(50):0.45±0.03 mg/L vs 0.15±0.03 mg/L,0.16±0.02 mg/L,P<0.01)的敏感性减低;流式细胞仪检测结果显示,转染细胞内阿霉素的蓄积减少(50.39±2.09 mg/L vs 94.99±4.07 mg/L,88.06±2.67 mg/L,P<0.01).结论:Ro 60变异体1真核表达载体转染SGC7901细胞后,显示多药耐药特性.
AIM： To investigate the possible role of Ro 60 variant 1 in multi-drug resistance （MDR） of gastric cancer.
METHODS： Ro 60 variant 1 encoding gene was cloned using RT-PCR method. Ro 60 variant 1 sense eukaryotic expression vector was constructed using DNA recombination technique and transfected into SGC7901 cells with Lipofectamine^TM 2000. Drug sensitivity was detected by MTT assay. IC50 values of gastric cancer cells for chemotherapy drugs were calculated. Intracellular accumulation of adriamycin in gastric cancer cells was measured by sorting fluorescence-activated cells.
RESULTS： The expression level of Ro 60 variant 1 in SGC7901 cells was increased after transfection with sense genes. In vitro drug sensitivity assay showed that the sensitivity of SGC7901 cells transfected with Ro 60 variant 1 genes was significantly decreased compared with SGC7901 and SGC7901-pcDNA3.1 cells on vincristine （IC50：2.87 ± 0.10 mg/L vs 0.47 ± 0.07 mg/L, 0.63 ± 0.08 mg/L, P 〈 0.01）, 5-FU（IC50： 3.89 ± 0.12 mg/L vs 0.59 ± 0.17 mg/L, 0.92 ± 0.12 mg/L, P〈0.01）, mitomycin （IC50：1.02 ± 0.06 mg/L vs 0.50 ± 0.04 mg/L, 0.73 ± 0.09 mg/L, P 〈 0.05）, cisplatin （IC50：1.15 ± 0.06 mg/L vs 0.46 ± 0.04 mg/L, 0.52 ± 0.05 mg/L, P 〈 0.01） and adriamycin （IC50：0.45 ± 0.03 mg/L vs 0.15 ± 0.03 mg/L, 0.16 ± 0.02 mg/L, P 〈 0.01）. Flow cytometry revealed that accumulation of adriamycin in SGC7901 cells transfected with Ro 60 variant 1 gene was decreased （50.39 ± 2.09 mg/L vs 94.99 ± 4.07 mg/L, 88.06 ± 2.67 mg/L, P 〈 0.01）, when compared with SGC7901 and SGC7901- pcDNA3.1 cells.
CONCLUSION： SGC7901 cells transfected with Ro 60 variant 1 sense genes exhibit MDR. Ro 60 variant 1 might play a certain role in MDR of gastric cancer.
出处
《世界华人消化杂志》
CAS
北大核心
2008年第8期814-818,共5页
World Chinese Journal of Digestology
作者简介
韩全利，2003年第四军医大学博士，主要从事胃癌多药耐药的研究.通讯作者：韩全利，100036，北京市海淀区阜成路30号，中国人民解放军空军总医院干部病房三区．hanquanli@yahoo.com．cn电话：010-66928118-6132 传真：010-66928132
