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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 observe the effect of new antisense oligonucleotide MBO-asGCS on the expression of glucosylceramide synthase (GCS) and P-glycoprotein (P-gp) in breast cancer cells. METHODS: Real-time polymerase chain reaction (PCR) was used to detect MCF-7-AdrR in breast cancer cells treated with MBO-asGCS. Western blot analysis of GCS and P-gp protein expression in MCF-7-AdrR and transformed cells was performed. TUNEL assay was used to detect MBO-asGCS. Apoptosis in treated MCF-7-AdrR. RESULTS: The expression of GCS and P-glycoprotein in MCF-7-AdrR/GCS overexpressed in drug-resistant strains and GCS was characteristically increased, whereas the expression of GCS and P-glycoprotein in asGCS-transformed cells MCF-7-AdrR/asGCS was increased. Significantly decreased; GCS gene expression in MCF-7-AdrR treated with MBO-asGCS was downregulated by 63.6%, and apoptotic cells were significantly increased (P<0.01). CONCLUSION: There may be a relationship between GCS and P-glycoprotein in the resistance mechanism of breast cancer cells. The elevated GCS can promote the metabolism of ceramide and increase the expression of P-glycoprotein, resulting in drug resistance. Antisense oligonucleotides Nucleotides can inhibit GCS expression, down-regulate P-glycoprotein and promote apoptosis.