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Using Northern and dot blot analysis, we investigated the expression of c-myc, c-fos abd erbB and regulation of oncogenes by TPA in BIU-87 cells a human bladder transitional cell carcinoma line We found expression of these oncogenes and induc-tion of these genes by TPA.We also found high expression of c-myc, c-fos, erbB, N-ras in human bladder cancer tissues.In contrast, only low level expression of N-ras was detected in normal bladder tissues.These data suggest that the expression of some oncogenes can be modulated... Using Northern and dot blot analysis, we investigated the expression of c-myc, c-fos abd erbB and regulation of oncogenes by TPA in BIU-87 cells a human bladder transitional cell carcinoma line We found expression of these oncogenes and induc-tion of these genes by TPA.We also found high expression of c-myc, c-fos, erbB, N-ras in human bladder cancer tissues.In contrast, only low level expression of N-ras was detected in normal bladder tissues.These data suggest that the expression of some oncogenes can be modulated in a protein kinase C dependent manner and high level expression of some oncogenes are responsible for the abnormal growth in bladder carcinoma. 本实验应用Northern和斑点印渍杂交技术探测了人膀胱癌细胞株中c-myc、c-fos、erbB等癌基因的表达,以及TPA对这些癌基因表达的调控,发现BIU-87细胞有这些癌基因的表达,并能被TPA所增强,同时也发现人膀胱癌组织有c-myc、c-fos、erbB、N-ras基因的高表达。提示蛋白激酶C的激活可以诱导某些癌基因的表达。多种癌基因的表达异常可能在膀胱癌中起重要作用。 The cDNA of hGM CSF was transfered into the human bladder cancer line BIU 87 with retroviral vector. PCR southern blot demonstrated the integration of transfered gene in the BIU cell genome. Biological activity of the hGM CSF secreted by the BIU cell was confirmed. DNA analysis by flowcytomerty showed no influence on growth of the BIU cells. Flourenscent immunoassay revealed no induction of HLA ABC, DR, DQ expression on the surface of the gene transfered BIU cells. The gene transfected BIU cells were irradiated... The cDNA of hGM CSF was transfered into the human bladder cancer line BIU 87 with retroviral vector. PCR southern blot demonstrated the integration of transfered gene in the BIU cell genome. Biological activity of the hGM CSF secreted by the BIU cell was confirmed. DNA analysis by flowcytomerty showed no influence on growth of the BIU cells. Flourenscent immunoassay revealed no induction of HLA ABC, DR, DQ expression on the surface of the gene transfered BIU cells. The gene transfected BIU cells were irradiated with 6000 rad X ray but they could secret hGM CSF for at least 2 weeks, although the growth of them was abrogated. These suggested that we could develope the bladder cancer vaccine secreting hGM CSF for clinical trials in future. 采用逆转录病毒载体将hGM-CSF基因导入人膀胱癌细胞株BIU-87细胞中,建立转基因细胞株BIU/GM。经PCR-SouthernBlot杂交证实GM-CSF基因已整合到BIU/GM细胞基因组中。经流式细胞仪细胞DNA周期分析表明,转基因前后BIU-87细胞增殖状态无变化。免疫荧光检测发现GM-CSF基因导入及表达不能促进BIU-87细胞表面HLA-ABC、DR、DQ抗原的表达。转基因细胞经60Gy/sX线灭活后,丧失增殖能力,但能维持一定水平的GM-CSF分泌活性达2周。本文为进行转基因瘤苗的深入研究奠定了初步基础。 A human bladder cancer cells line resistant to doxorubicin, BIU-87/ADM,has been established in vitro by exposing BIU-87 parent cell to progressively higher concentrations of the drug over a period of 8 months. The cell line has been characterized in terms of growth kinetics, morphology, cross-resistance to other anticancerous agents, pharmacokinetics of ADM and expression of p-glycoprotein(P-gp)which is closely related to the MDR phenotype, The BIU-87/ADM cell was 6.3 times more resistant to doxorubicin than... A human bladder cancer cells line resistant to doxorubicin, BIU-87/ADM,has been established in vitro by exposing BIU-87 parent cell to progressively higher concentrations of the drug over a period of 8 months. The cell line has been characterized in terms of growth kinetics, morphology, cross-resistance to other anticancerous agents, pharmacokinetics of ADM and expression of p-glycoprotein(P-gp)which is closely related to the MDR phenotype, The BIU-87/ADM cell was 6.3 times more resistant to doxorubicin than the BIU-87 parent cells. BIU-87/ADM exhibited cross-resistance to doxorubicin derivatives(epirubicin, daunorubicin),vincristine and etoposide, but not to cisplatin and mitomycin C, Compared to the parent cells, the resistant cells have a slower growth rate and lower confluent density. Unlike the BIU-87 parent cells, about 75% of the BIU 87/ADM cells showed a positive reaction with monoclonal antibody against P-gp,JSB-1. Intracellular drug accumulation studies with fluorescence spectrometry indicated that the resistance exhibited by the BIU-87/ADM line was mainly caused by an increased active efflux, These results suggest that MDR is an important phenomenon in bladder cancer and that more than one pathway of MDR may be present in human bladder cancer cell lines. BIU-87/ADM may be a useful model for the development of new chemotherapeutic strategies in overcoming drug-resistance in the treatment of bladder cancer. 以人膀胱癌细胞株BIU-87为亲本,经递增阿霉素(ADM)剂量的方法,历时8个月,建立了一株耐药亚株BIU-87/ADM.此细胞株对ADM的耐受程度较亲本细胞提高了6.3倍,对柔红霉素(DNR)、表阿霉素、长春新碱和鬼臼乙叉甙有明显的交叉耐药性,但对顺铂、丝裂霉素无交叉耐药性。与亲本细胞相比,耐药亚株生长慢,倍增期延长,汇合密度低,异形性明显,有巨细胞形成。进一步研究表明,耐药亚株对DNR蓄积减少;免疫化学显示,75%的耐药细胞P-gp过表达。因而耐药细胞内DNR低聚集主要是P-gp膜泵功能增强介导产生细胞内药物外溢增多所致,是其产生耐药性的主要原因。但并非所有BIU-87/ADM均表达P-gp,其他耐药机理的共存是有可能的。BIU-87/ADM及其亲本细胞为寻求逆转人膀胱癌耐药的新型化疗增敏剂或综合化疗方案提供了良好的实验模型。
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