摘要
目的 探讨CacyBP在胃癌多药耐药机制中的作用。方法 构建CacyBP正义核酸真核表达载体pcDNA3.1/hCacyBP +,以脂质体将其导入胃癌药敏细胞SGC790 1。以RT PCR检测hCacyBPmRNA水平的变化 ;以MTT检测SGC790 1及转染细胞对ADR的药物敏感性 ;以流式细胞仪 (FCM)检测SGC790 1及转染细胞内ADR的蓄积浓度及细胞周期。结果 转染pcDNA3.1/hCacyBP +的SGC790 1,其hCacyBPmRNA表达水平显著高于空载体转染及未转染之SGC790 1,且细胞生存率亦有所增高。未转染之SGC790 1及分别转染空载体或pcDNA3.1/hCacyBP +之SGC790 1细胞 ,平均ADR蓄积浓度分别为 5 .6 4 ,5 .4 9和 5 .17。与未转染之SGC790 1相比 ,转染pcDNA3.1/hCacyBP +和空载体的SGC790 1细胞G1期减少 ,G2 期及S期增高。
Objective To study the effect of human calcyclin binding protein (CacyBP) encoding gene on the development of multiple drug resistance in gastric cancer. Methods hCacyBP sense nucleic acid eukaryotic expression vector (pcDNA3.1/hCacyBP+) was constructed and then transfected steadily into the gastric cancer drug sensitive cell (SGC7901) mediated by Lipofectamine TM 2000. RT PCR was used to measure the CacyBP mRNA expression level. MTT was used to measure the adriamycin (ADR) sensitivity of SGC7901 and SGC7901 after transfection. FCM was used to measure the average ADR accumulation concentration and cell cycle of SGC7901 and SGC7901 after transfection. Results The hCacyBP mRNA expression level of SGC7901 transfected with pcDNA3.1/hCacyBP+ was higher than SGC7901 transfected with pcDNA3.1 or SGC7901, with the higher survival rate in the former. The average ADR accumulation concentration in SGC7901 and SGC7901 transfected with pcDNA3.1 or pcDNA3.1/hCacyBP+ was 5.64, 5.49 and 5.17, respectively. The G 1 phase cell proportion of SGC7901 transfected with pcDNA3.1/hCacyBP+ or pcDNA3.1 was reduced slightly but G 2 and S phases increased slightly as compared with SGC7901.Conclusion Calcyclin binding protein may play a certain role in gastric cancer drug resistance. [
出处
《中华肿瘤杂志》
CAS
CSCD
北大核心
2002年第5期426-429,共4页
Chinese Journal of Oncology
基金
国家自然科学基金资助项目 (3 0 0 3 0 14 03 0 0 2 40 0 2 )
关键词
CacyBP
胃癌
多药耐药
聚合酶链反应
基因转染
流式细胞仪
Stomach neoplasms
Calcyclin binding protein
Polymerase Chain reaction
Gene transfer
Flow cytometry