Respiration characteristics of mitochondria in parental and giant transformed cells of the murine Nemeth—Kellner lymphoma

Abstract

Respiration characteristics of mitochondria of the parental and giant cells of murine NK/Ly (Nemeth-Kellner lymphoma) were studied. The giant cell-enriched ascites were obtained by serial intraperitoneal injections of vinblastine in tumour-bearing mice. Ascites containing >70% giant cells were used. Their diameter of was over 17 μm (~2800 μm(3)), while the diameter of the parental cells was 12.7 μm (1100 μm(3)). The respiration rate of mitochondria in situ was measured by oxygen consumption in intact and digitonin-permeabilized NK/Ly cells. Endogenous respiration of intact giant NK/Ly cells was three times higher compared to the parental ones, roughly in agreement with the volume change. The giant NK/Ly cells were far more resistant to permeabilization with digitonin than the parental cells, as shown by Trypan Blue and LDH (lactate dehydrogenase) release tests. After digitonin permeabilization, oxygen consumption was reduced to a minimal level (0.06 ng atom O/(s × 106 cells) in both types of cells. Addition of α-ketoglutarate or succinate to the incubation medium increased oxygen consumption in the parental cells by 46 and 164% respectively. In the giant NK/Ly cells, the corresponding increases were 164 and 276%. Addition of ADP to α-ketoglutarate- or succinate-supplemented medium further stimulated oxygen consumption of the permeabilized NK/Ly cells; however, the effect of ADP was more pronounced in the giant cells. In addition, indices of respiratory control were significantly higher in the giant cells. Oligomycin suppressed considerably the respiration of the intact giant cells but had a much weaker effect on parental cells. Thus, giant NK/Ly cells possess much higher respiration rates and show tighter coupling between the respiration and oxidative phosphorylation compared with parental cells.

Influence of adenosine diphosphate on respiration of rat pancreatic acinar cells mitochondria in situ

“…Нашу увагу привернув метод дослідження дихання мітохондрій у пермеабілізованих клітинах, або за умов in situ, апробований на м'язових клітинах [14,19], гепатоцитах [1] і клітинах лімфоми [11]. Цей підхід є одним із способів поєднати переваги дослідження дихання ізольованих мітохондрій та цілісних клітин.…”

“…1,а,б). Як і в попередніх дослідженнях на інших тканинах [1,11], ФАД-залежний субстрат сукцинат суттєво інтенсифікував споживання кисню до 0,34 ± 0,04 нмоль О 2 / (с . 10 6 клітин) (P ≤ 0,01, n = 5; див.…”

A comparison of paclitaxel and vinblastine induced giant cell formation in murine NK/Ly lymphoma

“…Calculation of giant cells in population was proposed for evaluation of efficiency of chemotherapy. After treatment of tumor cells with cytostatic drugs, the number of giant cells increased up to eight times [11,12]. X-irradiation caused formation of giant cells that were 200 times bigger than the parental ones [4].…”

“…Giant cell transformation is strongly linked with changes in cytoskeleton, namely in microtubule system [2,9,11]. Microtubules are highly dynamic structures which constantly get longer and shorter [8].…”

“…In both cases, this results in cell cycle arrest which can occur in G 0 -or G 2 /M-pha ses [13]. Thus, the mitotic arrest is a common cause of giant cell formation [11,14]. In this study, we used two known anti-tubulin agents possessing opposite effects towards tubulin, and also used as anticancer drugs.…”