摘要
目的:对Ro 60反义核酸在逆转胃癌细胞多药耐药中作用进行研究．方法:克隆Ro 60编码基因,构建Ro 60编码基因的反义真核表达载体,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:成功构建了Ro 60反义真核表达载体,应用脂质体介导法将其转导入SGC7901-VCR, Ro 60反义真核表达载体转染SGC7901-VCR细胞后,Ro 60的表达量明显下降,体外药物敏感性实验提示其对长春新碱、丝裂霉素、顺铂、阿霉素的敏感性增加,转染反义表达载体的SGC7901-VCR细胞与未转染和转染空载体的细胞相比,IC50值(mg/L)有显著的下降(7．66±0．45 vs 19．56±0．38,17．48±0．85;0．84±0．03 vs 1．62±0．06．1．80±0．03;0．51±0．03 vs 0．87±0．03．0．88±0．03;0．22±0．01 vs 0．52±0．02,0．43±0．03,均P<0．01),细胞内阿霉素蓄积有显著的增加(51．94±1．26 mg／L vs 36．27±0．98,37．01±0．91 mg／L,P<0．01)．结论:Ro 60反义真核表达载体转染SGC7901后能够抑制胃癌细胞的多药耐药表型．
AIM： To investigate the possible function of Ro 60 antisense nucleic acid in multidrug resistant cell line of gastric cancer.
METHODS： Ro 60 antisense eukaryotic expression vector was constructed using DNA recombination technique, then transfected into SGC7901-VCR by LipofectamineTM2000. Drug sensitivity assay was performed using MTT assay, and ICs0 values of gastric cancer cells to chemotherapy drugs were calculated. The intracellular accumulation of adriamycin in gastric cancer cells was measured using fluorescenceactivated cell sorting
RESULTS： The expression level of Ro 60 in SGC7901-VCR cells was decreased after transfection with antisense genes. In vitro drug sensitivity assay show that SGC7901-VCR cells transfected with Ro 60 antisense genes showed significantly increased sensitivity to vincristine （IC50：7.66 ＋ 0.45 mg/L vs 19.56 ± 0.38, 17.48 ± 0.85 mg/L, P 〈 0.01）, mitomycin （IC50：0.84 ± 0.03 mg/L vs 1.62 ± 0.06, 1.80 ± 0.03 mg/L, P 〈 0.01）, cisplatin （ICs0： 0.51 ± 0.03 mg/L vs 0.87 ± 0.03, 0.88 ± 0.03 mg/L, P 〈 0.01） and adriamycin （ICs0：0.22 ± 0.01 mg/L vs 0.52 ± 0.02, 0.43 ± 0.03mg/L, P 〈 0.01）, as compared with SGC7901-VCR and SGC7901-VCR- pcDNA3.1 cells. As showed by flow cytometry, the intracellular accumulation of adriamycin in the cells transfected with Ro 60 antisense gene was markedly increased in comparison with that in SGC7901-VCR or SGC7901-VCR-pcDNA3.1 cells （51.94 ±1.26 mg/L vs 36.27 ± 0.98, 37.01 ± 0.91 rag/L, P 〈 0.01）.
CONCLUSION： After transfected into multidrug resistant cell line of gastric cancer, Ro 60 antisense nucleic acid can inhibit the multidrug resistant phenotype of gastric cancer.
出处
《世界华人消化杂志》
CAS
北大核心
2006年第27期2668-2672,共5页
World Chinese Journal of Digestology
基金
国家自然科学基金资助项目
No.30371623
