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Expression and activity of classic protein kinase C in drug-resistant gastric cancer cell line
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- Author:
- No author available
- Journal Title:
- JOURNAL OF THE FOURTH MILITARY MEDICAL UNIVERSITY
- Issue:
- 15
- DOI:
- 10.3321/j.issn:1000-2790.2001.15.005
- Key Word:
- 蛋白激酶C;同工酶;胃癌;多药耐药
Abstract： 目的观察传统型蛋白激酶C(cPKCs)在胃癌耐药细胞系SGC7901/VCR1.0、耐药亚系SGC7901/VCR0.3,SGC7901/VCR0.7及药敏细胞系SGC7901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9584.17, 9421.06, 8534.64, 8088.79,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SGC7901/VCR的多药耐药中起重要作用且具有同工酶特异性.
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