葡萄糖神经酰胺合成酶基因在人耐多柔比星乳腺癌细胞株MCF-7／ADR中的表达及意义  

Expression of GCS gene in human doxorubicin-resistant MCF-7/ADR cells and its significance

作　　者：胡萍萍[1] 陈同钰[1] 田波[2] 罗光华[1] 魏江[2] 

机构地区：[1]苏州大学附属第三医院病理科,江苏常州213003 [2]苏州大学附属第三医院综合实验室,江苏常州213003

出　　处：《中国癌症杂志》2008年第4期267-270,共4页China Oncology

摘　　要：背景与目的:多药耐药是肿瘤化疗失败的主要原因,近年来神经酰胺的糖基化水平增高与多药耐药的关系引起了广泛关注。本研究旨在探讨葡萄糖神经酰胺合成酶(GCS)基因在人乳腺癌细胞株MCF-7和人耐多柔比星(阿霉素)乳腺癌细胞株MCF-7/ADR中的表达及意义。方法:采用四氮唑盐(MTT)法检测多柔比星对MCF-7/ADR和MCF-7的抑制率和IC50,用浓度为5、10、20μmol/L的GCS抑制剂D,L-threo-1-phenyl-2-decanoyl-amino-3-morpholino-1-propanol(PDMP)预处理MCF-7/ADR24、48h后检测抑制率和IC50;应用实时荧光定量PCR(real time PCR)检测MCF-7、MCF-7/ADR以及PDMP预处理后MCF-7/ADR中GCS mRNA的表达。结果:MCF-7/ADR对MCF-7的耐药倍数为22.69倍,PDMP作用后多柔比星对MCF-7/ADR的抑制率升高,IC50从(19.0±0.5)μg/ml下降到(5.6±0.6)μg/ml。MCF-7/ADR中GCS mRNA的表达高于MCF-7(P<0.01),PDMP使MCF-7/ADR中GCS mRNA的表达水平GCSN(48±17)下降到(21±4)。结论:GCS可能参与乳腺癌的发生过程,并在MCF-7/ADR多药耐药中起重要作用。Background and purpose： Multidrug resistance（MDR） is considered to be the main obstacle to chemotherapy of malignant tumors, and the relationship between glucosyleeramide synthase and MDR has caught great attention recently. In this study, we investigated the expression of glucosylceramide synthase （GCS） gene in human MCF-7/ADR （doxorubicin-resistant） breast cancer cells and the parent MCF-7 cells and explored its significance. Methods： Exposing MCF-7/ADR to various concentrations（5, 10, 20 μmol/L） of GCS inbibitonr D, L-threo-1-phenyl-2-decanoyl-amino-3-morpholino -1- propanol（PDMP） for different time（24, 48 hr）, MTT was used to detect the drug resistance of MCF-7/ADR cells, the inhibition ratio and IC50 of MCF-7/ADR cells to doxorubicin was investigated as to whether they were pretreated by PDMP or not; Real time RT-PCR was adopted to evaluate the expression of GCS mRNA in them. Results： The drug resistance of MCF-7/ADR was 22.69 times, GCS mRNA expressing in both cells and their expression was significantly increased compared with MCF-7 ceils by real time RT-PCR. PDMP exposure enhanced doxorubicin sensitivity, it decreased the IC50 of MCF-7/ADR ceils from （ 19.0±0.5） μg/ml to （5.6±0.6）μg/ml, and the expression of GCSN decreased from （48±17） to （21±4） after pretreatment with it. Conclusions： GCS, expressing in human breast carcinoma cells, may take part in the carcinogenesis and development of breast carcinoma and play an important role in the drug resistance of it.

关 键 词：葡萄糖神经酰胺合成酶 乳腺癌细胞株  多药耐药 

分 类 号：R737.9[医药卫生—肿瘤]