Spaggiare S, Wallach M J, Tupper J T
J Cell Physiol. 1976 Nov;89(3):403-16. doi: 10.1002/jcp.1040890306.
The components of unidirectional K influx and efflux have been investigated in the 3T3 cell and the SV40 transformed 3T3 cell in expontntial and stationary growth phase. Over the cell densities used for transport experiments the 3T3 cell goes from exponential growth to density dependent inhibition of growth (4 X 10(4) to 4 X 10(5) cell cm-2) whereas the SV40 3T3 maintains exponential or near exponential growth (4 X 10(4) to 1 X 10(6) cell cm-2). In agreement with previous observations, volume per cell and mg protein per cell decrease with increasing cell density. Thus, transport measurements have been expressed on a per volume basis. Total unidirectional K influx and efflux in the 3T3 cell is approximately double that of the SV40 3T3 cell at all cell densities investigated. Both cell types have similar volumes initially and show similar decreases with increasing cell density. Thus, in this clone of the 3T3 cell SV40 transformation specifically decreases unidirectional K flux. The magnitude of the total K flux does not change substantially for either cell line during transition from sparse to dense cultures. However, the components of the K transport undergo distinct changes. Both cell lines possess a ouabain sensitive component of K influx, presumably representing the active inward K pump. Both also possess components of K influx and efflux sensitive to furosemide. The data suggest this component represents a one-for-one K exchange mechanism. The fraction of K influx mediated by the ouabain sensitive component is reduced to one half its value when exponential versus density inhibited 3T3 cells are compared (63% versus 31% of total influx). No comparable drop occurs in the SV40 3T3 cell at equivalent cell densities (64% versus 56% of total influx). Thus, the pump mediated component of K influx would appear to be correlated with growth. In contrast, the furosemide sensitive component represents approximately 20% of the total unidirectional K influx and efflux in both cell lines in sparse culture. At high cell densities, where growth inhibition occurs in the 3T3 cell but not the SV40 3T3, the furosemide sensitive component doubles in both cell lines. Thus, the apparent K-K exchange mechanism is density dependent rather than growth dependent.
在指数生长期和静止期的3T3细胞以及SV40转化的3T3细胞中,对单向钾离子流入和流出的组成部分进行了研究。在用于转运实验的细胞密度范围内,3T3细胞从指数生长转变为密度依赖性生长抑制(从4×10⁴到4×10⁵个细胞/cm²),而SV40 3T3细胞保持指数或接近指数生长(从4×10⁴到1×10⁶个细胞/cm²)。与先前的观察结果一致,每个细胞的体积和每个细胞的蛋白质毫克数随着细胞密度的增加而减少。因此,转运测量结果以每体积为基础表示。在所有研究的细胞密度下,3T3细胞的总单向钾离子流入和流出量约为SV40 3T3细胞的两倍。两种细胞类型最初的体积相似,并且随着细胞密度的增加显示出相似的减少。因此,在这个3T3细胞克隆中,SV40转化特异性地降低了单向钾离子通量。对于两种细胞系,在从稀疏培养到致密培养的转变过程中,总钾离子通量的大小没有实质性变化。然而,钾离子转运的组成部分发生了明显变化。两种细胞系都具有对哇巴因敏感的钾离子流入成分,推测代表主动内向钾离子泵。两者还具有对速尿敏感的钾离子流入和流出成分。数据表明该成分代表一对一的钾离子交换机制。当比较指数生长的3T3细胞和密度抑制的3T3细胞时,由哇巴因敏感成分介导的钾离子流入比例降至其值的一半(分别占总流入量的63%和31%)。在相同细胞密度下,SV40 3T3细胞没有出现类似的下降(分别占总流入量的64%和56%)。因此,钾离子流入的泵介导成分似乎与生长相关。相反,速尿敏感成分在稀疏培养的两种细胞系中约占总单向钾离子流入和流出量的20%。在高细胞密度下,3T3细胞出现生长抑制而SV40 3T3细胞没有,此时两种细胞系中速尿敏感成分均增加一倍。因此,明显的钾离子-钾离子交换机制是密度依赖性的,而不是生长依赖性的。