摘要
目的 观察传统型蛋白激酶C(cPKCs)在胃癌耐药细胞系SGC790 1/VCR1.0、耐药亚系SGC790 1/VCR0 .3,SGC790 1/VCR0 .7及药敏细胞系SGC790 1表达及活性的变化 ,以及对细胞内药物浓度的影响 .方法 用免疫荧光化学及蛋白印迹方法观察传统型蛋白激酶C在胃癌耐药细胞系及药敏细胞系的表达 ;竞争蛋白结合法测定PKC的活性 ;应用流式细胞仪观察细胞内药物浓度的变化 .结果 PKCα,PKCβⅠ ,PKCβⅡ 及PKCγ在胃癌耐药细胞系及药敏细胞系均有表达 ,PKCα在耐药细胞表达呈强阳性 ,在药敏细胞表达呈阳性 ;PKCβⅠ 及PKCβⅡ 在耐药细胞及药敏细胞表达均为阳性 ;PKCγ在耐药细胞及药敏细胞表达均为强阳性 .免疫蛋白印迹实验证实 ,随耐药指数的增加 ,PKCα表达呈逐渐增高的趋势 ,薄层扫描蛋白印迹带的吸光度 (A)分别为 95 84.17,942 1.0 6 ,85 34 .6 4,80 88.79,而PKCβⅠ ,PKCβⅡ 及PKCγ在耐药细胞系、耐药亚系及其药敏细胞系表达无明显变化 .PKC活性检测结果提示 ,随耐药指数的增加 ,PKC活性呈逐渐增高的趋势 ,以细胞质及细胞核的PKC活性增高为主 .结论 传统型蛋白激酶C在维持胃癌耐药细胞系SGC790
AIM To observe the expression and activity of classic protein kinase C in gastric cancer cell SGC7901 and its drug resistant sublines SGC7901/VCR slected by vincristine of different concentration (0.3, 0.7 and 1.0 μg·mL -1 ). To explore the effect of classic protein kinase C on drug concentration within gastric cancer cells. METHODS The expression of classic protein kinase C in gastric cancer cells was determined by immuno fluorescent method and Western blot. The PKC activity was determined by competed protein binding method. The drug concentration within cells was determined by FACS. RESULTS Both SGC7901 and SGC7901/VCR cells exhibited positive staining of PKCα, PKCβ Ⅰ, PKCβ Ⅱ and PKCγ. The staining of PKCα in SGC7901/VCR was much stronger than that in SGC7901. There were no differences in the expression of PKCβ Ⅰ, PKCβ Ⅱ and PKCγ between SGC7901 and SGC7901/VCR. By using Western blot, continuous increasing expression of PKCα in SGC7901/VCR was demonstrated along with the increasing resistant index, as well as no differences between SGC7901 and its drug resistant sublines. Results of activity assay showed continuous increasing activity of PKC in gastric cancer cells along with increasing resistant index. CONCLUSION Classic protein kinase C plays an important role in multidrug resistance of gastric cancer cells with isoenzyme specificity.
出处
《第四军医大学学报》
北大核心
2001年第15期1358-1361,共4页
Journal of the Fourth Military Medical University
基金
国家自然科学基金资助课题 (39880 0 0 7)
国家杰出青年基金资助课题 (3952 50 2 0 )
关键词
蛋白激酶C
同工酶
胃癌
多药耐药
protein kinase C
isoenzyme
gastric cancer
multidrug resistance
