人视网膜母细胞瘤耐药细胞系SO-Rb50/CBP的建立及耐药相关基因表达检测  

Establishment of a multidrug resistance cell line SO-Rb50 /CBP of human retinoblastoma and expression analysis of drug resistance associated genes

作　　者：杨玉霞[1] 孟永[1,2] 周立军[1,3] 李永平[1] 郑健樑[1] 张文忻[1] 林健贤[1] 张平[1] 

机构地区：[1]中山大学中山眼科中心中山大学眼科学国家重点实验室,广州510060 [2]无锡市第三人民医院眼科 [3]湖南省儿童医院眼科

出　　处：《中华临床医师杂志（电子版）》2010年第11期85-90,共6页Chinese Journal of Clinicians(Electronic Edition)

基　　金：国家自然科学基金(30371515)

摘　　要：目的用卡铂诱导人视网膜母细胞瘤SO-Rb50成为耐药细胞SO-Rb50/CBP,检测其生物学特性,并探讨其耐药机制。方法以人视网膜母细胞瘤SO-Rb50为亲本细胞,采用卡铂高浓度间歇诱导法诱导多药耐药细胞SO-Rb50/CBP。MTT法测定其药物敏感性及生长曲线、倍增时间;分别用免疫细胞化学、流式细胞术对MRP、GST-π蛋白表达进行检测,用RT-PCR检测耐药相关基因MRP、GST-π的表达。结果耐药细胞SO-Rb50/CBP的耐药指数为15.7。亲本细胞与耐药细胞群体倍增时间分别为44.32h和50.9h。SO-Rb50/CBP对CBP、DDP、VCR、MMC、VP-16和5-FU的耐药指数分别为15.7、6.89、5.68、4.58、2.28和1.88。免疫细胞化学SO-Rb50/CBP的GST-π和MRP表达的阳性率分别为96.19%和90.21%,亲本细胞两种蛋白的阳性率分别为20.12%和30.48%。SO-Rb50/CBP的MRP及GST-π的mRNA表达水平比SO-Rb50明显增高。结论诱导出耐卡铂的视网膜母细胞瘤细胞SO-Rb50/CBP呈高度耐药,且存在多药耐药现象。其耐药现象的产生,可能与GST-π和MRP耐药蛋白表达的增加有关。Objective To establish a carboplatin-resistance human retinoblastoma cell line SO-Rb50 /CBP and study its biological characteristics and the probable drug-resistance mechanism. Methods First,SO-Rb50 /CBP was induced by intermittently administration of high-concentration of Carboplatin（ CBP）. Then MTT assay was used to detect drug sensitivity of the two cell lines. Then reverse transcripts polymerase chain reaction （ RT-PCR） was employed to detect the transcription of MRP and GST-π genes in cells of SO-Rb50 and SO-Rb50/CBP respectively. Immunohistochemical technique and flow cytometry were used to detect the proteins of MRP and GST-π of the above cells. Results The drug-resistance index of SO-Rb50 /CBP cell line to CBP was 15. 7. The population doubling time of SO-Rb50 and SO-Rb50 /CBP were 44. 32 h and 50. 9 h respectively. The drug-resistance indexs of SO-Rb50 /CBP to DDP,VCR,MMC,VP-16 and 5-FU were 6. 89,5. 68,4. 58,2. 28 and 1. 88 respectively. The positive rate of GST-πand MRP in SO-Rb50 /CBP cells were 96. 19% and 90. 21% respectively by immunocytochemistry,while in SO-Rb50 cells were 20. 12% and 30. 48% respectively. The mRNA level of MRP and GST-πwere higher in SO-Rb50 /CBP cells than that in SO-Rb50 cells. Conclusions The established carboplatin-resistance human retinoblastoma cell line SO-Rb50 /CBP showed resistance to several other drugs such as MMC,DDP,VCR,VP-16. The drug-resistance mechanism of SO-Rb50 /CBP cells maybe due to high expression of MRP and GST-π genes.

关 键 词：视网膜母细胞瘤 抗药性 肿瘤 卡铂 细胞系 肿瘤 

分 类 号：R739.7[医药卫生—肿瘤]