Alkalinization of the lysosomes is correlated with ras transformation of murine and human fibroblasts.
Alkalinization of the lysosomes is correlated with ras transformation of murine and human fibroblasts.
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DOI：
10.1016/s0021-9258(19)34037-2
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发表时间：
1990-03
期刊：
影响因子：
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通讯作者：
Lian-Wei Jiang;V. Maher;J. Mccormick;Melvin Schindler
中科院分区：
文献类型：
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作者：
Lian-Wei Jiang;V. Maher;J. Mccormick;Melvin Schindler
通过测定488和457 nm激发的液相胞嘧啶化荧光素共轭右旋糖酐（FITC-dextrans）在530 nm处的荧光发射强度，监测培养成纤维细胞溶酶体内腔室的pH。遵循Ohkuma和Poole （Ohkuma， S., and Poole， B., 1978）的程序。学会科学。美国（1978）75,3327-3331)，建立了fitc -葡聚糖的荧光发射强度与pH之间的关系。这种相关性被用于测定一系列成纤维细胞培养的溶酶体内表观pH （pHapp）。未转化的小鼠3T3成纤维细胞和无限寿命的人成纤维细胞株MSU-1.1的酶内体平均pHapp值为5.0。分别转染v-Ki-ras或T24 H-ras致癌基因转化的MSU-1.1细胞和Kirsten小鼠肉瘤病毒转化的3T3成纤维细胞的恶性率为6.1。这些测量结果表明，ras转化导致溶酶体pH值的显著扰动。
The pH of the intralysosomal compartment of fibroblasts in culture was monitored by measuring the fluorescence emission intensity at 530 nm of fluid phase pinocytosed fluorescein-conjugated dextrans (FITC-dextrans) excited at 488 and 457 nm. Following the procedure of Ohkuma and Poole (Ohkuma, S., and Poole, B. (1978) Proc. Natl. Acad. Sci. U. S. A. (1978) 75, 3327-3331), a relationship was established between the fluorescence emission intensity of the FITC-dextrans and pH. This correlation was used to determine the intralysosomal apparent pH (pHapp) of a series of fibroblast cultures. The mean intralysosomal pHapp values of nontransformed mouse 3T3 fibroblasts and an infinite life-span human fibroblast cell strain, designated MSU-1.1, was 5.0. In distinction that of 3T3 fibroblasts transformed to the malignant state by Kirsten murine sarcoma virus and MSU-1.1 cells transformed by transfection of the v-Ki-ras or T24 H-ras oncogene was 6.1. These measurements suggest that ras transformation results in a significant perturbation of lysosomal pH.