目的研究鞘磷脂合酶2(sphingomyelin synthase 2,SMS2)缺损对人结肠癌细胞(colon cancer cell,Caco-2)中药物转运体P-糖蛋白(P-glycoprotein,P-gp)及多药耐药相关蛋白2(multidrug resistance-associated protein 2,MRP2)的表达与功能的...目的研究鞘磷脂合酶2(sphingomyelin synthase 2,SMS2)缺损对人结肠癌细胞(colon cancer cell,Caco-2)中药物转运体P-糖蛋白(P-glycoprotein,P-gp)及多药耐药相关蛋白2(multidrug resistance-associated protein 2,MRP2)的表达与功能的影响。方法 SMS2特异性siRNA转染至Caco-2细胞。采用RT-PCR和real-time PCR检测SMS2-特异性siRNA的干扰效率;采用Western blot法检测siRNA转染后P-gp和MRP2蛋白表达的变化;采用胞内蓄积试验,通过HPLC检测P-gp及MRP2专一性底物罗丹明123及普伐他汀的蓄积含量,分析下调SMS2水平对Caco-2细胞中P-gp及MRP2功能的影响。结果应用siRNA下调SMS2水平后,P-gp和MRP2的蛋白质水平均显著下调;胞内蓄积实验结果显示罗丹明123及普伐他汀的蓄积量明显增加,说明P-gp和MRP2的外排功能减弱。结论SMS2基因表达下调对Caco-2细胞中P-gp和MRP2的表达以及功能均有显著影响。展开更多
鼠源抗人乳腺癌多药耐药相关蛋白1(multidrug resistance-associated protein 1,MRP1/ABCC1)噬菌体Fab抗体库的构建首先是利用乳腺癌组织匀浆液作为免疫原免疫BALB/c小鼠,成功免疫后提取小鼠脾组织的总RNA逆转录为cDNA。然后设计小鼠Ig...鼠源抗人乳腺癌多药耐药相关蛋白1(multidrug resistance-associated protein 1,MRP1/ABCC1)噬菌体Fab抗体库的构建首先是利用乳腺癌组织匀浆液作为免疫原免疫BALB/c小鼠,成功免疫后提取小鼠脾组织的总RNA逆转录为cDNA。然后设计小鼠IgG各亚型的特异性Fab引物,PCR扩增Fd和L基因并依次连接到噬菌体载体pComb3中。重组体pComb3-Fab转化至E.coli XL1-Blue菌中,最后经辅助噬菌体VCSM13超感染,成功构建噬菌体Fab抗体库。以化学合成的MRP1/ABCC1膜表位10肽为抗原进行3轮淘选、富集,对淘选后获得的各级噬菌体抗体库进行ELISA检测和鉴定。成功获得的抗MRP1/ABCC1-10肽三级噬菌体Fab抗体库将为抗人乳腺癌MRP1/ABCC1 Fab抗体的制备提供保障。展开更多
Objective: To investigate the relationship between the expression of multidrug resistance-associated protein (MRP) and clinicopathological factors and prognosis. Methods: The expression of MRP in 62 cases with non-sma...Objective: To investigate the relationship between the expression of multidrug resistance-associated protein (MRP) and clinicopathological factors and prognosis. Methods: The expression of MRP in 62 cases with non-small cell lung cancer (NSCLC) was detected using immunohistochemistry method. The expression of MRP in 30 cases of NSCLC and corresponding normal lung tissues were detected using immunohistochemistry and Western Blot. Results: this study of tumor tissues confirmed the plasma membrane and/or cytoplasm locations of MRP. There was apparent difference between normal lung tissues and NSCLC in MRP. The survival analysis of 62 NSCLC showed that the mean survival time of the patients with negative MRP expression was 69.8117.41 months and that of patients with positive MRP expression, 25.384.46 months. Log-rank test suggested that the difference between them was significant (P=0.0156). It was also found that in squamous cell lung cancer the statistically significant difference between the mean survival time of patients with positive MRP expression and those with negative MRP expression (P=0.0153). Multivariate Cox model analysis suggested that the survival time was significantly related to expression of MRP (P=0.035) and lymphatic metastasis (P=0.038). Conclusion: MRP expression in NSCLC is significantly higher compared with normal lung tissues. The mean survival time of patients with negative MRP was relative longer and expression of MRP was an independent factor for prognosis.展开更多
原发性肝癌(primary of liver)是我国常见的恶性肿瘤之一.手术切除率相对较低,术后复发率较高,许多肿瘤常规化疗效果差,预后不良,而肿瘤多药耐药性(MDR)则是肿瘤化疗失败的关键因素.多药耐药相关蛋白(multidrug associated protein,MRP...原发性肝癌(primary of liver)是我国常见的恶性肿瘤之一.手术切除率相对较低,术后复发率较高,许多肿瘤常规化疗效果差,预后不良,而肿瘤多药耐药性(MDR)则是肿瘤化疗失败的关键因素.多药耐药相关蛋白(multidrug associated protein,MRP)是一种介导多药耐药性的跨膜转运蛋白,能减少细胞内药物聚积,或改变药物在细胞内分布,从而影响化疗疗效及患者生存.主要针对MRP的结构和功能、在正常组织和肝癌细胞中的表达,以及多药耐药相关蛋白MRP的耐药机制和耐药性逆转等因素进行综述,以为临床提高肝癌的综合治疗水平提供理论基础.展开更多
Objective: To investigate the effects of neoadjuvant chemotherapy on the expression of drug resistance genes, multidrug resistance-1 (MDR1) and multidrug resistance-associated protein (MRP), in patients with primary b...Objective: To investigate the effects of neoadjuvant chemotherapy on the expression of drug resistance genes, multidrug resistance-1 (MDR1) and multidrug resistance-associated protein (MRP), in patients with primary breast cancer. Methods: MDR1 and MRP expression were detected by semi-quantitative RT-PCR in 20 patients with primary breast cancer, before and after chemotherapy. Results: Before chemotherapy, MDR1 and MRP expression can be detected in 15 cases (75%) and 18 cases (90%) respectively. After chemotherapy, expression of MDR1 is not significantly different from that before chemotherapy, but expression of MRP is significantly different from that before chemotherapy. Conclusion: Expression of drug resistance gene MRP, but not MDR1, is enhanced in patients with primary breast cancer submitted to neoadjuvant chemotherapy.展开更多
文摘Objective: To investigate the relationship between the expression of multidrug resistance-associated protein (MRP) and clinicopathological factors and prognosis. Methods: The expression of MRP in 62 cases with non-small cell lung cancer (NSCLC) was detected using immunohistochemistry method. The expression of MRP in 30 cases of NSCLC and corresponding normal lung tissues were detected using immunohistochemistry and Western Blot. Results: this study of tumor tissues confirmed the plasma membrane and/or cytoplasm locations of MRP. There was apparent difference between normal lung tissues and NSCLC in MRP. The survival analysis of 62 NSCLC showed that the mean survival time of the patients with negative MRP expression was 69.8117.41 months and that of patients with positive MRP expression, 25.384.46 months. Log-rank test suggested that the difference between them was significant (P=0.0156). It was also found that in squamous cell lung cancer the statistically significant difference between the mean survival time of patients with positive MRP expression and those with negative MRP expression (P=0.0153). Multivariate Cox model analysis suggested that the survival time was significantly related to expression of MRP (P=0.035) and lymphatic metastasis (P=0.038). Conclusion: MRP expression in NSCLC is significantly higher compared with normal lung tissues. The mean survival time of patients with negative MRP was relative longer and expression of MRP was an independent factor for prognosis.
文摘原发性肝癌(primary of liver)是我国常见的恶性肿瘤之一.手术切除率相对较低,术后复发率较高,许多肿瘤常规化疗效果差,预后不良,而肿瘤多药耐药性(MDR)则是肿瘤化疗失败的关键因素.多药耐药相关蛋白(multidrug associated protein,MRP)是一种介导多药耐药性的跨膜转运蛋白,能减少细胞内药物聚积,或改变药物在细胞内分布,从而影响化疗疗效及患者生存.主要针对MRP的结构和功能、在正常组织和肝癌细胞中的表达,以及多药耐药相关蛋白MRP的耐药机制和耐药性逆转等因素进行综述,以为临床提高肝癌的综合治疗水平提供理论基础.
文摘Objective: To investigate the effects of neoadjuvant chemotherapy on the expression of drug resistance genes, multidrug resistance-1 (MDR1) and multidrug resistance-associated protein (MRP), in patients with primary breast cancer. Methods: MDR1 and MRP expression were detected by semi-quantitative RT-PCR in 20 patients with primary breast cancer, before and after chemotherapy. Results: Before chemotherapy, MDR1 and MRP expression can be detected in 15 cases (75%) and 18 cases (90%) respectively. After chemotherapy, expression of MDR1 is not significantly different from that before chemotherapy, but expression of MRP is significantly different from that before chemotherapy. Conclusion: Expression of drug resistance gene MRP, but not MDR1, is enhanced in patients with primary breast cancer submitted to neoadjuvant chemotherapy.
