Objective The clinical signilicance of exPression of multidrug resistance- associated protein (MRP) in gastric and renal carcinoma was investigated. Methods LSAB immunohistochemistry was performed to detect eopression...Objective The clinical signilicance of exPression of multidrug resistance- associated protein (MRP) in gastric and renal carcinoma was investigated. Methods LSAB immunohistochemistry was performed to detect eopression of MRP in the carcinoma tissues of 52 patients with gastric carcinoma and 20 cases with renal cell carcinoma. Results The positive expression rate of MRP was 38.5% (20/52) in gastric carcinoma tissues, and 60% (12/20) in renal carcinoma tissues. The expression of MRP both on cellular membrane and in cytoplasm was observed, but the expression in cytoplasm (thick granule) was more obvious. The positive expression rates of MRP in advanced gastric and renal carcinoma (Ⅲ orⅣ stage) were 60% (15/25) and 88.90% (8/9) reSPectively, which were higher than those in early lesion (Ⅰ or Ⅱ stage, 18.5% and 36.4% respectively). Furthermore, the patients with positive expression of MRP in gastric carcinoma tissues had shorter mean survival time and lower 5-year survival rate than that with negative eopression of MRP. Conclusion MRP plays an important role in the infiltration and metastasis of gastric and renal carcinoma and might contribute to the intrinsic drug - resistance in both carcinomas.展开更多
目的:观察环孢素A(CsA)处理急性单核细胞白血病细胞株THP-1后对柔红霉素(DNR)的耐药性及与多药耐药相关蛋白-1(MRP1)表达的关系,探讨CsA对THP-1细胞的化疗增敏作用及相关机制。方法:噻唑蓝(MTT)法、荧光分光光度法、共聚焦激光扫描显微...目的:观察环孢素A(CsA)处理急性单核细胞白血病细胞株THP-1后对柔红霉素(DNR)的耐药性及与多药耐药相关蛋白-1(MRP1)表达的关系,探讨CsA对THP-1细胞的化疗增敏作用及相关机制。方法:噻唑蓝(MTT)法、荧光分光光度法、共聚焦激光扫描显微镜(CLSM)法分别检测CsA处理前后50% THP-1细胞生长受抑的DNR浓度(IC50)、THP-1细胞内DNR含量及分布的变化。免疫细胞化学法、Western blot法检测CsA处理前后THP-1细胞MRP1蛋白表达量的变化。结果:CsA处理后THP-1细胞的IC50明显下降(1.5364±0.1751 vs 0.5588±0.0547,P<0.01),细胞内DNR含量明显增加(21.40±1.71 vs 131.96±16.45,P<0.01),且胞浆内分布更加均匀;同时MRP1蛋白表达量增高(0.2665±0.0042 vs 0.3169±0.0062,P<0.01)。结论:CsA可增加THP-1细胞对DNR的敏感性,降低其耐药性,并提示CsA对THP-1细胞耐药性的改变不是通过降低其MRP1蛋白表达量来实现的。展开更多
目的:研究甲氨蝶呤(methotrexate,MTX)耐药前后人大肠癌(结直肠癌)HT-29细胞中透明质酸蛋白聚糖连结蛋白1(hyaluronan and proteoglycan link protein 1,HAPLN1)表达的变化及其对MTX耐药性的影响,并探究可能的分子机制。方法:采用浓度...目的:研究甲氨蝶呤(methotrexate,MTX)耐药前后人大肠癌(结直肠癌)HT-29细胞中透明质酸蛋白聚糖连结蛋白1(hyaluronan and proteoglycan link protein 1,HAPLN1)表达的变化及其对MTX耐药性的影响,并探究可能的分子机制。方法:采用浓度梯度递增法构建耐药细胞株HT-29/MTX;RT-PCR检测HAPLN1和多药耐药相关蛋白2(multidrug resistance-associated protein 2,MRP2)的mRNA表达水平;采用脂质体介导法将人HAPLN1和MRP2基因的干扰质粒转染HT-29/MTX细胞并筛选出稳定表达的细胞系;采用CCK-8法检测细胞活力;流式细胞术检测细胞凋亡;采用Western blot检测HAPLN1、MRP2、IκB激酶(IκB kinase,IKK)α/β、p-IKKα/β(Ser176/Ser177)、p65和p-p65(Ser536)的蛋白水平。结果:HT-29/MTX细胞中HAPLN1和MRP2的mRNA和蛋白表达水平都显著高于其亲代HT-29细胞(P<0.05),耐药倍数高达463.756。抑制HAPLN1和MRP2基因表达使MTX对HT-29/MTX细胞的IC_(50)从15.304μmol/L分别降至6.119μmol/L和7.801μmol/L,逆转倍数分别为2.501和1.962,并增强MTX诱导的细胞凋亡(P<0.05)。敲减HAPLN1基因表达和使用IKK抑制剂IKK16均可下调IKKα/β和p65蛋白磷酸化水平以及MRP2蛋白表达水平(P<0.05),但IKK16未对HAPLN1蛋白表达产生影响(P>0.05)。结论:敲减HAPLN1基因表达可在体外增强人大肠癌耐药细胞株HT-29/MTX对MTX的敏感性,这可能与其抑制IKK/p65信号通路活化,继而下调MRP2基因表达有关。展开更多
进一步提高晚期鼻咽癌(nasopharyngeal carcinoma,NPC)患者生存率的主要障碍是较高的远处转移率,出现远处转移的主要原因之一是由P糖蛋白(P-glycoprotein,Pgp)和多药耐药相关蛋白-1(multidrug resistanceassociated protein 1,MRP1)介...进一步提高晚期鼻咽癌(nasopharyngeal carcinoma,NPC)患者生存率的主要障碍是较高的远处转移率,出现远处转移的主要原因之一是由P糖蛋白(P-glycoprotein,Pgp)和多药耐药相关蛋白-1(multidrug resistanceassociated protein 1,MRP1)介导的多药耐药现象(multidrug resistance,MDR),因此研究Pgp和MRP1引起的MDR对提高NPC的预后具有重要意义。本文就Pgp、MRP1的检测方法和逆转剂及其在NPC中的应用进行综述。展开更多
文摘Objective The clinical signilicance of exPression of multidrug resistance- associated protein (MRP) in gastric and renal carcinoma was investigated. Methods LSAB immunohistochemistry was performed to detect eopression of MRP in the carcinoma tissues of 52 patients with gastric carcinoma and 20 cases with renal cell carcinoma. Results The positive expression rate of MRP was 38.5% (20/52) in gastric carcinoma tissues, and 60% (12/20) in renal carcinoma tissues. The expression of MRP both on cellular membrane and in cytoplasm was observed, but the expression in cytoplasm (thick granule) was more obvious. The positive expression rates of MRP in advanced gastric and renal carcinoma (Ⅲ orⅣ stage) were 60% (15/25) and 88.90% (8/9) reSPectively, which were higher than those in early lesion (Ⅰ or Ⅱ stage, 18.5% and 36.4% respectively). Furthermore, the patients with positive expression of MRP in gastric carcinoma tissues had shorter mean survival time and lower 5-year survival rate than that with negative eopression of MRP. Conclusion MRP plays an important role in the infiltration and metastasis of gastric and renal carcinoma and might contribute to the intrinsic drug - resistance in both carcinomas.
文摘目的:观察环孢素A(CsA)处理急性单核细胞白血病细胞株THP-1后对柔红霉素(DNR)的耐药性及与多药耐药相关蛋白-1(MRP1)表达的关系,探讨CsA对THP-1细胞的化疗增敏作用及相关机制。方法:噻唑蓝(MTT)法、荧光分光光度法、共聚焦激光扫描显微镜(CLSM)法分别检测CsA处理前后50% THP-1细胞生长受抑的DNR浓度(IC50)、THP-1细胞内DNR含量及分布的变化。免疫细胞化学法、Western blot法检测CsA处理前后THP-1细胞MRP1蛋白表达量的变化。结果:CsA处理后THP-1细胞的IC50明显下降(1.5364±0.1751 vs 0.5588±0.0547,P<0.01),细胞内DNR含量明显增加(21.40±1.71 vs 131.96±16.45,P<0.01),且胞浆内分布更加均匀;同时MRP1蛋白表达量增高(0.2665±0.0042 vs 0.3169±0.0062,P<0.01)。结论:CsA可增加THP-1细胞对DNR的敏感性,降低其耐药性,并提示CsA对THP-1细胞耐药性的改变不是通过降低其MRP1蛋白表达量来实现的。
文摘目的:研究甲氨蝶呤(methotrexate,MTX)耐药前后人大肠癌(结直肠癌)HT-29细胞中透明质酸蛋白聚糖连结蛋白1(hyaluronan and proteoglycan link protein 1,HAPLN1)表达的变化及其对MTX耐药性的影响,并探究可能的分子机制。方法:采用浓度梯度递增法构建耐药细胞株HT-29/MTX;RT-PCR检测HAPLN1和多药耐药相关蛋白2(multidrug resistance-associated protein 2,MRP2)的mRNA表达水平;采用脂质体介导法将人HAPLN1和MRP2基因的干扰质粒转染HT-29/MTX细胞并筛选出稳定表达的细胞系;采用CCK-8法检测细胞活力;流式细胞术检测细胞凋亡;采用Western blot检测HAPLN1、MRP2、IκB激酶(IκB kinase,IKK)α/β、p-IKKα/β(Ser176/Ser177)、p65和p-p65(Ser536)的蛋白水平。结果:HT-29/MTX细胞中HAPLN1和MRP2的mRNA和蛋白表达水平都显著高于其亲代HT-29细胞(P<0.05),耐药倍数高达463.756。抑制HAPLN1和MRP2基因表达使MTX对HT-29/MTX细胞的IC_(50)从15.304μmol/L分别降至6.119μmol/L和7.801μmol/L,逆转倍数分别为2.501和1.962,并增强MTX诱导的细胞凋亡(P<0.05)。敲减HAPLN1基因表达和使用IKK抑制剂IKK16均可下调IKKα/β和p65蛋白磷酸化水平以及MRP2蛋白表达水平(P<0.05),但IKK16未对HAPLN1蛋白表达产生影响(P>0.05)。结论:敲减HAPLN1基因表达可在体外增强人大肠癌耐药细胞株HT-29/MTX对MTX的敏感性,这可能与其抑制IKK/p65信号通路活化,继而下调MRP2基因表达有关。
