葡萄糖神经酰胺合成酶基因在人胰腺癌细胞SW1990及其耐药亚株中的表达和意义

目的探讨葡萄糖神经酰胺合成酶（glucosylceramide synthase,GCS）基因在人胰腺癌细胞SW1990及其耐药亚株中的表达和意义。方法体外培养人胰腺癌细胞SW1990和其耐药亚株SW1990／FU、SW1990／ADM、SW1990／GEM,以细胞活性测定（CCK-8法）检测耐药亚株的耐药指数,相对定量荧光实时PCR（RT—PCR）法检测GCS mRNA的表达,免疫印迹法检测GCS蛋白表达水平。结果与亲本株相比,SW1990／FU、SW1990／ADM、SW1990／GEM的耐药指数分别为339．7、11．9、56．6。RT—PCR和免疫印迹检测结果显示亲本株和耐药亚株均有GCS mRNA和蛋白的表达,GCS mRNA在3株耐药亚株中的表达均比亲本株有增高趋势,且SW1990／ADM、SW1990／GEM的GCS蛋白表达水平较亲本株升高,差异有统计学意义（P〈0．05）。结论GCS基因在人胰腺癌细胞株SW1990和其3株耐药亚株中表达阳性,GCS蛋白在SW1990／ADM、SW1990／GEM中呈高表达,表明GCS可能参与胰腺癌耐药株细胞对ADM、GEM的耐药。

Expression and significance of GCS gene in human pancreatic cancer cell SW1990 and its drug-resistant sublines

Objective To study the expression and significance of GCS gene in human pancreatic cancer cell line SW1990 and its drug-resistant sublines.Methods SW1990 and its drug-resistant sublines, SW1990/FU,SW1990/ADM and SW1990/GEM were cultured in vitro.CCK-8(Cell Counting kit-8)was used to detect the drug resistance of the sublines.Relative quantitation of GCS mRNA expression was evaluated by real-time PCR and Western blot was adopted to evaluate the expression of GCS protein.Results The drug resistance indexes of SW1990/FU,SW1990/ADM and SW1990/GEM were 339.7,11.9 and 56.6,respectively.The GCS mRNA and protein were expressed in all SW1990 and its drug-resistant sublines.There was a higher expression of GCS mRNA in all the sublines and a significant difference of GCS protein expression was detected in SW1990/ADM and SW1990/GEM compared with SW1990.Conclusions GCS gene is expressed in SW1990 and its drug-resistance sublines.The high expression of GCS protein in SW1990/ADM and SWI990/GEM might be one reason of resistance to ADM and GEM in the sublines.