生长抑素调控细胞内药物浓度增强阿霉素对胆囊癌细胞的杀伤效能  

Somatostatin enhanced anti-tumor effect of doxorubicin on gallbladder caner cells through the regulation of intracellular drug concentration

作　　者：龚伟[1] 秦一雨[1] 李松岗[1] 全志伟[1] 李济宇[1] 庄鹏远[1] 万紫微[1] 

机构地区：[1]交通大学医学院附属新华医院普外科,上海200092

出　　处：《中华医学杂志》2010年第38期2718-2722,共5页National Medical Journal of China

摘　　要：目的 探讨生长抑素（SST）增强阿霉素（DOX）杀伤胆囊癌细胞株（GBC-SD）的作用机制.方法 以GBC-SD细胞为实验对象,按处理因素不同,共分为4组：SST组、DOX组、SST-DOX联合用药组和仅添加等量磷酸盐缓冲液（PBS）的对照组.根据我们以前的研究结果,药物作用浓度分别为SST（75 μg/ml）,DOX（5μg/ml）.药物作用0,6,12,24,36 h后,MTT法观察各组细胞活性;荧光分光光度法检测各组细胞内DOX浓度的变化;分别采用Real-time PCR与Western印迹检测药物作用后各时间点细胞多药耐药基因1（MDR1）mRNA表达及P-糖蛋白表达（P-gp）的变化情况.结果 单独应用SST没有明显抑制GBC-SD细胞生长的作用（P＞0.05）;药物作用12 h,与空白组比较,DOX组与SST＋DOX组细胞杀伤显著增多,但DOX与SST＋DOX组间差异无统计学意义;SST联合作用24 h可显著增强DOX对GBC-SD细胞的杀伤作用,与DOX组比较差异有统计学意义（P＜0.05）.与DOX比较,SST联合作用可升高细胞内DOX浓度,二者间差异以联合作用24 h后最为显著（P＜0.05）;SST作用可显著降低GBC-SD细胞MDR1 mRNA表达（P＜0.05）及其转录翻译产物P-gp蛋白表达（P＜0.05）.结论 SST通过降低GBC-SD细胞内MDR1基因和P-gp蛋白的表达水平,使GBC-SD细胞泵出DOX减少,提高了细胞内DOX浓度,增强DOX对GBC-SD细胞的杀伤作用.Objective To investigate the possible mechanisms by which Somatostatin （SST）enhances the anti-tumor effect of doxorubicin （DOX） on gallbladder cancer cells. Methods GBC-SD cells were grouped into 4 groups： SST-treated group, DOX-treated group, SST ＋ DOX co-treated group and control group. The concentrations of SST and DOX were 75 μg/ml and 5 μg/ml based on our previous studies. In control group, cells were cultivated with phosphate buffered saline （PBS）. In experimental groups, cells were cultivated with medium and the corresponding drugs. After drug treatment, cell viability was examined by MTT assay at 6, 12, 24 and 36 h respectively. Meanwhile, intracellular concentrations of doxorubicin in each group was determined by microspectrofluorimetry; Real-time polymerase chain reaction （RT-PCR） was used to determine the expressions of MDR1 mRNA in the cells at different time points and the expressions of P-gp protein, a product of MDR1 mRNA, were determined by Western blot analysis. Results SST did not exhibit significant inhibitory effect on the proliferation of GBC-SD cells as compared to that of control group （P 〉 0. 05）. SST ＋ DOX co-treatment group and DOX showed significantly inhibitory effect on the growth of GBC-SD cells at Hour 12 post-treatment. However no statistical difference was found between SST ＋ DOX and DOX groups. Interestingly, at Hour 24 post-treatment, SST ＋ DOX group showed more robust inhibitory effect on GBC-SD cells as compared to DOX alone group. Moreover, SST could significantly down-regulate the expressions of MDR1 mRNA and P-gp protein. SST could increase intracellular DOX concentration. And the difference of intracellular DOX concentration between SST ＋ DOX group and DOX group at Hour 24 was statistically significant. Conclusions In our experiment, SST decreases the expression of MDR1 mRNA and P-gp protein so as to reduce the efflux of DOX and elevate DOX concentrations in GBC-SD cells. This eventually leads to enhanced cytotoxic effects of DOX on GBC-

关 键 词：胆囊肿瘤 生长抑素 P糖蛋白 阿霉素 细胞内药物浓度 

分 类 号：R686[医药卫生—骨科学]