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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 investigate the biological characteristics of SO-Rb50 / SO-Rb50 induced by retinoblastoma SO-Rb50 by carboplatin and explore its mechanism of drug resistance. Methods Human retinoblastoma SO-Rb50 was used as the parent cell, and the multi-drug resistant cell line SO-Rb50 / CBP was induced by high concentration of carboplatin intermittently. The drug sensitivity, growth curve and doubling time were measured by MTT assay. The expression of MRP and GST-πprotein were detected by immunocytochemistry and flow cytometry respectively. The expression of MRP and GST-π expression. Results The resistant index of SO-Rb50 / CBP in drug-resistant cells was 15.7. The doubling time of parental cells and drug-resistant cell groups were 44.32h and 50.9h, respectively. The resistance index of SO-Rb50 / CBP to CBP, DDP, VCR, MMC, VP-16 and 5-FU were 15.7, 6.89, 5.68, 4.58, 2.28 and 1.88, respectively. The positive rates of GST-π and MRP expression in immunocytochemistry SO-Rb50 / CBP were 96.19% and 90.21%, respectively. The positive rates of the two proteins in parental cells were 20.12% and 30.48%, respectively. The mRNA expression levels of MRP and GST-π in SO-Rb50 / CBP were significantly higher than SO-Rb50. Conclusion The resistance to carboplatin-induced retinoblastoma cells SO-Rb50 / CBP is highly resistant and multi-drug resistant. The drug resistance phenomenon may be related to the increase of GST-π and MRP resistance protein expression.