摘要
目的 观察葡萄糖神经酰胺合酶(glucosylcerarnide synthase,GCS)基因在人口腔表皮样癌多药耐药细胞株KBV_(200)的表达及其与肿瘤多药耐药性(MDR)的关系。方法 采用RT-PCR分析KBV_(200)及其亲本KB细胞株GCS基因表达的差异,运用MTT法分析KBV_(200)经糖脂合成酶抑制剂苯基棕榈酰胺吗啡丙醇(DL-PPMP)及钙离子通道阻滞剂异搏定处理前后细胞MDR的变化,应用RT-PCR技术检测KBV_(200)细胞耐药逆转后GCS及mdrl基因的表达。结果 KBV_(200)细胞GCS和mdrl基因的表达明显强于KB细胞,而KB细胞mdrl基因表达为阴性。5～25μmol/L DL-PPMP均可抑制KBV_(200)GCS mRNA表达,25 μmol/L时抑制强度最大;10 μmol/L异搏定即可抑制KBV_(200)GCS mRNA表达,15 μmol/L时抑制作用明显。DL-PPMP和异搏定均可抑制mdrl基因的表达,KBV_(200)经25μmol/L DL-PPMP作用48h后细胞内mdrl表达为阴性。结论 异搏定和DL-PPMP对GCS和marl基因表达的抑制调节呈浓度依赖性,它们可通过抑制GCS和mdrl基因表达,逆转KBV_(200)对长春新碱的耐药。KBV_(200)GCS基因的表达与MDR存在正相关,GCS基因可能在肿瘤的多药耐药过程中起着重要作用。
Objective To investigate the relationship between the expression of glucosylceramide synthase (GCS) mRNA in vincristine-resistant KBV_(200) human cancer cell line and multidrug resistance (MDR) of the cancer cells. Methods Reverse transcriptional polymerase chain reaction (RT-PCR) was employed to analyze the differential expression of GCS mRNA between KBV_(200) and KB cell lines and the changes in the mRNA expressions of GCS and mdrl gene in KBV_(200) cells after reversion of MDR. The effects of de-phenyl-z-palmaitoylamino-3-morpholine-1-propanol (DL-PPMP) and verapamil in reversing MDR of the cells were evaluated by MTT assay. Results KBV_(200) cells exhibited significantly increased expressions of GCS and mdrl gene, whereas mdrl gene failed to be detected in the parental KB cells. DL-PPMP within the concentrations ranging from 5 to 25 μmol/L could inhibit the expression of GCS gene, with the maximum inhibition achieved at 25 μmol/L. Verapamil at the concentration of 10 μmol/L was already sufficient to induce inhibition of GCS expression in KVB_(200) cells, which was more manifest with the concentration of 15 μmol/L. DL-PPMP and verapamil were found to inhibit mdrl gene expression in KBV_(200) cells at the mRNA level, and complete inhibition occurred after a 48-hour DL-PPMP treatment at 25 μmol/L. Conclusion The inhibition of GCS and mdrl gene expressions is positively correlated with the concentrations of DL-PPMP and verapamil, which can reverse MDR by inhibiting synthesis of GCS and mdrl gene, indicating the positive correlation between the expression of GCS gene and MDR in KBV_(200) cells. GCS gene might play an importantrole in MDR during tumor progression.
出处
《第一军医大学学报》
CSCD
北大核心
2004年第7期779-783,共5页
Journal of First Military Medical University
基金
广东省自然科学基金(001041)~~
