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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 SWl990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,GCSmRNA 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。
Objective To investigate the expression and significance of glucosylceramide synthase (GCS) gene in human pancreatic cancer cell line SW1990 and its resistant subline. Methods Human pancreatic cancer cell line SW1990 / SW1990 / SW, SW1990 / ADM and SW1990 / GEM were cultured in vitro. The drug resistance index of resistant subline was detected by CCK-8 assay. The relative quantitative fluorescence The expression of GCS mRNA was detected by PCR (RT-PCR), and the expression of GCS protein was detected by Western blotting. Results Compared with the parent strain, the resistance index of SW1990 / FU, SW1990 / ADM and SW1990 / GEM were 339.7, 11.9 and 56.6 respectively. The results of RT-PCR and Western blotting showed that the expression of GCS mRNA and protein were both in the parental and drug resistant sub-strains. The expression of GCSmRNA in the three resistant sub-strains was higher than that in the parental strain, and SW1990 / ADM and SW1990 / GEM GCS protein expression levels higher than the parent strain, the difference was statistically significant (P <0.05). Conclusions The GCS gene was expressed in human pancreatic cancer cell line SW1990 and its 3 resistant sub-strains. GCS protein was highly expressed in SW1990 / ADM and SW1990 / GEM, indicating that GCS may be involved in the proliferation of ADM, GEM resistance.