Russo M A, Galeotti T, van Rossum G D
Cancer Res. 1976 Nov;36(11 Pt 1):4160-74.
The changes in water and electrolyte content of slices of Morris hepatoma 3924A induced by various conditions of incubation have been compared with the ultrastructural appearance of the tissue. Incubation at 1 degrees led to an increase of water, Na+, and Cl- content and to a loss of K+. There was simultaneous increase in size of the cells and intercellular spaces, loss of junctional complexes, increase in the number of microvilli, and fragmentation and dilation of the cytocavitary network. Subsequent incubation at 38 degrees in oxygenated medium led to a substantial reversal of all of these changes of composition and structure, which was well advanced within 10 min and largely complete by 60 min. The presence of 20 mM glucose in the medium somewhat enhanced the degree of recovery. A reduction of cell volume and intercellular spaces was evident both from the electron microscopic observations and measurements of the volume of inulin distribution. The presence of ouabain inhibited the net accumulation of K+ and much of the Na+ extrusion, but permitted about 50% of the net extrusion of water (accompanied by Na+ and Cl-) and had little effect on the ultrastructural recovery. The presence of glucose increased the resistance of volume and structural recovery of ouabain without releasing the inhibition of K+ accumulation. A marked feature of the recovering tissues was the Golgi apparatus, which assumed an appearance suggestive of increased activity when water extrusion was active. In slices using only endogenous substrate, cyanide and (to a lesser extent) oligomycin greatly inhibited the recovery of volume and structure. The presence of glucose permitted some recovery in the presence of cyanide. The control of cell volume in hepatoma 3924A appears to involve two separate components of water transport, one of which is sensitive, and one insensitive to ouabain. The ouabain-insensitive component appears to be especially related to the recovery of cell ultrastructure after incubation at 1 degrees, to be more sensitive to paucity of adenosine 5'-triphosphate, and to proceed by secretion of water, Na+, and Cl- into vesicles that fuse with the Golgi apparatus. This mechanism may be related to that for bile secretion in normal liver. The ouabain-sensitive component of water transport is a function of the mechanism for the coupled transport of Na+ and K+.
将经各种孵育条件诱导的Morris肝癌3924A切片的水和电解质含量变化与组织的超微结构外观进行了比较。在1℃孵育导致水、Na⁺和Cl⁻含量增加以及K⁺丢失。细胞和细胞间隙大小同时增加，连接复合体丢失，微绒毛数量增加，细胞腔网络碎片化和扩张。随后在38℃的含氧培养基中孵育导致所有这些组成和结构变化的显著逆转，在10分钟内进展良好，60分钟时基本完成。培养基中存在20 mM葡萄糖在一定程度上增强了恢复程度。从电子显微镜观察和菊粉分布体积测量都明显看出细胞体积和细胞间隙减小。哇巴因的存在抑制了K⁺的净积累和大部分Na⁺的排出，但允许约50%的水（伴随Na⁺和Cl⁻）净排出，并且对超微结构恢复影响很小。葡萄糖的存在增加了哇巴因处理后体积和结构恢复的抵抗力，而不解除对K⁺积累的抑制。恢复组织的一个显著特征是高尔基体，当水排出活跃时，其外观显示出活性增加。在仅使用内源性底物的切片中，氰化物和（程度较轻的）寡霉素极大地抑制了体积和结构的恢复。葡萄糖的存在使氰化物存在时仍有一定程度的恢复。3924A肝癌细胞体积的控制似乎涉及水运输的两个独立成分，其中一个对哇巴因敏感，另一个不敏感。对哇巴因不敏感的成分似乎特别与在1℃孵育后细胞超微结构的恢复有关，对三磷酸腺苷缺乏更敏感，并通过将水、Na⁺和Cl⁻分泌到与高尔基体融合的小泡中来进行。这种机制可能与正常肝脏中胆汁分泌的机制有关。水运输中对哇巴因敏感的成分是Na⁺和K⁺协同运输机制的一个功能。
