靶向葡萄糖神经酰胺合成酶逆转乳腺癌多药耐药的研究  

Targeting glucosylceramide synthase reverses multi-drug resistance in breast carcinoma cells

作　　者：孙妍琳[1] 周庚寅[1] 赵胜梅[2] 牟坤[1] 张翠娟[1] 张晓芳[1] 相磊[1] 

机构地区：[1]山东大学医学院病理学教研室,山东济南250012 [2]山东大学齐鲁医院检验科,山东济南250012

出　　处：《山东大学学报（医学版）》2005年第10期877-880,884,共5页Journal of Shandong University:Health Sciences

基　　金：国家自然科学基金资助课题(30300124);山东省科技厅重点支持课题(022130112)。

摘　　要：目的:探讨靶向葡萄糖神经酰胺合成酶(glucosylceramide synthase,GCS)对乳腺癌多药耐药的逆转作用及机制。方法:GCS反义寡核苷酸(GCSASODN)转染人耐药乳腺癌细胞MCF-7/AdrR,RT-PCR检测细胞GCS和多药耐药基因1(MDR1)mRNA的表达,免疫细胞化学染色检测细胞GCS蛋白和P-糖蛋白(P-gp)的表达,流式细胞仪分析细胞周期和凋亡。Caspase-3活性测定法检测Caspase-3活性改变。结果:GCSASODN转染人耐药乳腺癌细胞MCF-7/AdrR后,GCS和MDR1mRNA的表达水平分别为0.3、0.7,GCS蛋白和P-gp表达阳性率分别为23%、33%,GCS和MDR1的mRNA和蛋白水平较对照组有显著性差异(P<0.05)。转染后细胞周期改变不明显,但细胞凋亡增加为13.7±4.1,Caspase-3活性升高为0.0725±0.0003,与对照组差异有显著性(P<0.01)。结论:靶向GCS通过直接抑制GCS活性,并间接下调MDR1mRNA和蛋白表达,激活Caspase-3活性,诱导耐药细胞凋亡增加,有效逆转乳腺癌多药耐药。Objective： To study the effect and mechanism of targeting glucosylceramide synthase （GCS） on multi-drug resistance in breast carcinoma. Methods： GCS antisense oligodeoxynucleotide （GCS ASODN）was used to transfect human drug-resistant breast carcinoma cells MCF-7/AdrR, the mRNA expression of GCS and MDR was studied by using RT-PCR analysis, the protein expression of GCS and P-gp was studied by immunocytochemistry. Flow cytemetry analysis was used to detect the ratio of apoptosis and cell cycle. The alteration of Caspase-3 activity was assessed by Caspase-3 activity assess method. Results： After MCF-7/AdrR cells transfected with GCS ASODN, GCS and MDR1 expression was 0.3 and 0.7 respectively, the positive rate of GCS and P-gp expression was 23% and 33% respectively. There was significant difference of GCS and MDR1 expression compared with the control （P〈0.05）. Cell cycle was not altered, but the apoptosis of cells increased to 13.7±4.1, the activity of Caspase-3 increased to 0.0725±0.0003, （P〈0.01）. Conclusion： Targeting GCS can reverse multi-drug resistance in breast carcinoma efficiently by inhibiting GCS directly ,down-regulating MDRlmRNA and protein expression indirectly, activating Caspase-3 and inducing apoptosis of drug-resistant cells.

关 键 词：葡萄糖神经酰胺合成酶 乳腺肿瘤 抗药性 多药 

分 类 号：R737.9[医药卫生—肿瘤]