摘要
目的:观察新型反义寡核苷酸MBO-asGCS对乳腺癌细胞中葡萄糖神经酰胺合酶(GCS)与P-糖蛋白(P-gp)表达的影响。方法:实时PCR检测MBO-asGCS处理的耐药型乳腺癌细胞MCF-7-AdrR,蛋白质印迹分析MCF-7-AdrR及其转化细胞中GCS与P-gp的蛋白表达,TUNEL法检测MBO-asGCS处理的MCF-7-AdrR中的细胞凋亡情况。结果:在耐药株和GCS过表达的MCF-7-AdrR/GCS中GCS和P-糖蛋白表达特征性上升,而在asGCS转化细胞MCF-7-AdrR/asGCS中GCS和P-糖蛋白表达明显下降;MBO-asGCS处理的MCF-7-AdrR中GCS基因表达下调63.6%,凋亡细胞显著增加(P<0.01)。结论:乳腺癌细胞的耐药机制中可能存在GCS和P-糖蛋白的某种关联,升高的GCS可促进神经酰胺的代谢并增强P-糖蛋白的表达,产生耐药现象,反义寡核苷酸可抑制GCS表达,下调P-糖蛋白并促进凋亡。
Objective： To investigate the effect on expression of glucosylceramide synthase（GCS） and P-glycoprotein in drug-resistant breast cancer cells under a new oligonucleotide（MBO-asGCS） treatment.Methods： GCS mRNA expression was analyzed by realtime-PCR on MCF-7-AdrR cell treated with MBO-asGCS,expression of GCS and P-glycoprotein were assessed by western-blot analysis,apoptosis cells were labeled by terminal deoxynucleotide transferase-mediated dUTP nick end labeling（TUNEL）.Results： Both of GCS and P-glycoprotein overexpressed in the drug-resistant MCF-7-AdrR and MCF-7-AdrR/GCS,whereas reduced expression of GCS and P-glycoprotein were observed in the MCF-7-AdrR/asGCS transfected by antisense GCS.Furthermore,MBO-asGCS dramatically suppressed GCS mRNA（63.6%,P0.01） but increased apoptotic cells greatly in MCF-7-AdrR（P0.01）.Conclusion： GCS may associate with P-glycoprotein in the drug-resistant mechanism,which increased GCS enhanced ceramide metabolism and upregulated P-glycoprotein resulted in drug-resistant phenomenon;antisense oligonucleotide inhibited expression GCS and P-glycoprotein,further induced the cell apoptosis.
出处
《江苏大学学报（医学版）》
CAS
2010年第6期461-464,共4页
Journal of Jiangsu University：Medicine Edition
基金
国家自然科学基金资助项目(30770666)
