Sheedlo H J, Starosta-Rubinstein S, Desmond T J, Siegel G J
J Neuroimmunol. 1987 Jun;15(2):173-84. doi: 10.1016/0165-5728(87)90091-9.
The content and distribution of the membrane-bound enzyme (Na+ + K+)-ATPase in a rat cerebral C6 glioma was determined by immunocytochemistry, immunoblots and enzyme assay. In the C6 glioma cell culture (Na+ + K+)-ATPase activity was about 20% of (Mg2+ + Na+ + K+)-ATPase activity. However, (Mg2+ + Na+ + K+)-ATPase activity in the cerebral C6 gliomas was very close to Mg2+ baseline and not significantly increased by Na+ and K+. As shown by immunoblotting, (Na+ + K+)-ATPase catalytic subunit was detected in excised samples of control cerebrum and as a trace in the intracerebral portions of C6 glioma but not at all in the extracranial portions of C6 glioma or in C6 glioma cell culture. (Na+ + K+)-ATPase was not detected immunocytochemically in paraffin sections of the extracranial or intracerebral portions of rat cerebral C6 glioma. The absence of staining for (Na+ + K+)-ATPase clearly demarcated projections of glioma within normal brain. These results suggest that C6 glioma has little if any expression of (Na+ + K+)-ATPase in vitro or in vivo. The small amount of enzyme epitope in the intracerebral portions represents contamination by normal cerebrum in the extracts.
通过免疫细胞化学、免疫印迹和酶活性测定,确定了大鼠脑C6胶质瘤中膜结合酶(Na⁺ + K⁺)-ATP酶的含量和分布。在C6胶质瘤细胞培养物中,(Na⁺ + K⁺)-ATP酶活性约为(Mg²⁺ + Na⁺ + K⁺)-ATP酶活性的20%。然而,脑C6胶质瘤中的(Mg²⁺ + Na⁺ + K⁺)-ATP酶活性非常接近Mg²⁺基线,且未因Na⁺和K⁺而显著增加。免疫印迹显示,在对照大脑的切除样本中检测到了(Na⁺ + K⁺)-ATP酶催化亚基,在C6胶质瘤的脑内部分有微量存在,但在C6胶质瘤的颅外部分或C6胶质瘤细胞培养物中完全未检测到。在大鼠脑C6胶质瘤颅外或脑内部分的石蜡切片中,未通过免疫细胞化学检测到(Na⁺ + K⁺)-ATP酶。(Na⁺ + K⁺)-ATP酶染色缺失清楚地划分了正常脑内胶质瘤的投射。这些结果表明,C6胶质瘤在体外或体内几乎没有(Na⁺ + K⁺)-ATP酶的表达。脑内部分少量的酶表位代表提取物中正常大脑的污染。