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目的探讨葡萄糖神经酰胺合成酶(glucosylceramidesynthase,GCS)基因在人乳腺癌细胞的表达及意义。方法体外培养人耐阿霉素乳腺癌细胞MCF7/ADR和人敏感乳腺癌细胞MCF7,MTT法检测MCF7/ADR细胞耐药倍数,RT PCR法检测两种细胞GCSmRNA的表达,免疫细胞化学法检测GCS蛋白表达水平。结果MCF7/ADR细胞耐药倍数为53倍,RT PCR检测结果显示两种细胞均有GCSmRNA表达,且MCF7/ADR细胞GCSmRNA表达水平较MCF7细胞有显著升高(P<0.05),免疫细胞化学结果显示二者GCS蛋白均有表达且表达水平有显著差异(P<0.05)。结论GCS基因在乳腺癌细胞中表达,GCS对于乳腺癌的发生发展具有重要作用,GCS在耐药乳腺癌细胞的高表达证明GCS与肿瘤耐药密切相关,为肿瘤耐药的逆转治疗提供了新的方向。
Objective To investigate the expression of glucosylceramides synthase (GCS) gene in human breast cancer cells and its significance. Methods Human adriamycin-resistant breast cancer cells MCF7 / ADR and human sensitive breast cancer cells MCF7 were cultured in vitro. The drug resistance of MCF7 / ADR cells was detected by MTT assay. The expression of GCS mRNA was detected by RT-PCR and GCS Protein expression levels. Results The multidrug resistance of MCF7 / ADR cells was 53-fold. RT-PCR results showed that both cells had GCS mRNA expression, and the expression of GCS mRNA in MCF7 / ADR cells was significantly higher than that in MCF7 cells (P <0.05). Immunocytochemistry The results showed that both GCS proteins were expressed and the expression levels were significantly different (P <0.05). Conclusion GCS gene expression in breast cancer cells, GCS plays an important role in the occurrence and development of breast cancer. The high expression of GCS in resistant breast cancer cells proved that GCS is closely related to tumor drug resistance, providing a new drug for the reversal of drug resistance The direction of