Biological response of multicellular emt6 spheroids to exogenous lactate

Bourrat-Floeck, B.; Groebe, Karlfried; Mueller‐Klieser, Wolfgang

doi:10.1002/ijc.2910470528

Cited by 40 publications

(35 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Fig. S1) showed a similar growth behavior to that described for human carcinoma HepG2, human glioblastoma SNB19, mouse mammary EMT6 spheroids (Bourrat-Floeck et al, 1991;Hauptmann et al, 1998;Kelm et al, 2003), and T47-D and Hek293 spheroids (Gallardo-Pérez JC and Maldonado V, unpublished work). In all these models the maximal diameters reached were around 1 mm.…”

Section: Morphology and Growth Of Mctssupporting

confidence: 67%

Energy metabolism transition in multi‐cellular human tumor spheroids

Rodrı́guez-Enrı́quez

Gallardo‐Pérez

Avilés‐Salas

et al. 2008

Journal Cellular Physiology

124

101

View full text Add to dashboard Cite

It is thought that glycolysis is the predominant energy pathway in cancer, particularly in solid and poorly vascularized tumors where hypoxic regions develop. To evaluate whether glycolysis does effectively predominate for ATP supply and to identify the underlying biochemical mechanisms, the glycolytic and oxidative phosphorylation (OxPhos) fluxes, ATP/ADP ratio, phosphorylation potential, and expression and activity of relevant energy metabolism enzymes were determined in multi-cellular tumor spheroids, as a model of human solid tumors. In HeLa and Hek293 young-spheroids, the OxPhos flux and cytochrome c oxidase protein content and activity were similar to those observed in monolayer cultured cells, whereas the glycolytic flux increased two- to fourfold; the contribution of OxPhos to ATP supply was 60%. In contrast, in old-spheroids, OxPhos, ATP content, ATP/ADP ratio, and phosphorylation potential diminished 50-70%, as well as the activity (88%) and content (3 times) of cytochrome c oxidase. Glycolysis and hexokinase increased significantly (both, 4 times); consequently glycolysis was the predominant pathway for ATP supply (80%). These changes were associated with an increase (3.3 times) in the HIF-1alpha content. After chronic exposure, both oxidative and glycolytic inhibitors blocked spheroid growth, although the glycolytic inhibitors, 2-deoxyglucose and gossypol (IC(50) of 15-17 nM), were more potent than the mitochondrial inhibitors, casiopeina II-gly, laherradurin, and rhodamine 123 (IC(50) > 100 nM). These results suggest that glycolysis and OxPhos might be considered as metabolic targets to diminish cellular proliferation in poorly vascularized, hypoxic solid tumors.

show abstract

Section: Morphology and Growth Of Mctssupporting

confidence: 67%

Energy metabolism transition in multi‐cellular human tumor spheroids

Rodrı́guez-Enrı́quez

Gallardo‐Pérez

Avilés‐Salas

et al. 2008

Journal Cellular Physiology

124

101

View full text Add to dashboard Cite

show abstract

“…The interactions observed between these nutrients suggested that EMT6 cells in spheroids can adapt their metabolic rates substantially to different supply conditions (Mueller-Klieser et al 1986). as well as to the concentration of lactate (Bourrat-Floeck et al 1991). On this basis.…”

Section: Resultsmentioning

confidence: 85%

Variable presence of hypoxia in M006 human glioma spheroids and in spheroids and xenografts of clonally derived sublines

Franko¹,

Parliament²,

Allalunis-Turner³

et al. 1998

Br J Cancer

View full text Add to dashboard Cite

Summary Recently we reported the variable presence of hypoxia adjacent to necrosis in human glioma lines grown as subcutaneous tumours in severe combined immunodeficient (SCID) mice. To assess the basis for this observation. we examined the pattem of oxygenation in M006 and M006XLo glioma spheroids. We found a wide range of binding of [3H] aExduding two spheroids with a mixture of high and low grains ( Figure 3B).completely at each feeding. The number of spheroids w-as reduced at each feeding to stabilize the rate of consumption of nutrients. The first spheroids reached diameters of 0.8-1.2 mm. at x-hich they w-ere eliaible for experiments. w ithin 4-5 eeks. M006XLo sublineIn an effort to denrve a subline that w-as adapted to grow-th at lowambient oxy gen. the M006XLo subline x as established from small M006 spheroids that had been exposed continuously to 0.6%7 oxygen for 13 days. beainning 1 week after initiation in air. The spheroids were disaggregated wxith 0.25%c trypsin. and the cells w ere groxx n in monolayer culture for four passages (5 w eeks). then injected subcutaneously in SCID mice. A tumour x-as disaggregated to yield a new line designated M006XLo. w hich w as propagated in monolayer culture. Cells from the third passage were stored in liquid nitrogen. and each line x-as discarded after 3 months in culture and replaced from frozen stock.Clonally derived sublines (Franko et al. 1987). The desired oxygen level was obtained by a series of partial evacuations. each of which was followed by refilling of the chambers with 95% nitroaenlr5% carbon dioxide. Metabolic actixation of misonidazole during the degrassing procedure w-as minimized by precooling the medium and the chambers to 0°C and maintainingr this temperature during, the 30-to 40-min degassing process. The incubation period was 3 h for the spinner flask. and an additional 30 min for the aluminum chambers. w-hich is the time required for the medium in the dishes to return to 37' in the cabinet used for incubation. The oxygen tension in each chamber was measured at the end of the incubation period. as described previously (Franko et al. 1987). AutoradiographyThe spheroids were fixed in formalin and embedded in wax. and 4-gm serial sections w-ere taken throughout the spheroids. Slides w-ith sections near the centres of the majority of the spheroids were

show abstract

“…Similar proliferation-related changes occur in spheroids with a decrease in the overall oxygen uptake of spheroids during the cultivation period (Freyer et al, 1984;Freyer and Sutherland, 1985;Mueller-Klieser et al, 1986a;Bourrat-Floeck et al, 1991). In contrast, the respiratory activity within these spheroids seems to be rather uniform despite a nonuniform distribution of proliferating and quiescent cells (Mueller-Klieser, 1984;MuellerKlieser et al, 1986a;Bourrat-Floeck et al, 1991).…”

mentioning

confidence: 88%

“…The former parameter resulted in a mean value ( 5 SEM) of (3.8 ? 0.25) x m10,/cm3/s for EMTG spheroids with necrosis in a n experimental series published elsewhere (Bourrat-Floeck et al, 1991). An autoradiographic picture and the thymidine labeling index LI a t various depths into the spheroid is shown in a and b, respectively, of Figure 5.…”

Section: Multicellular Emtg Spheroidsmentioning

confidence: 98%

Proliferation‐associated oxygen consumption and morphology of tumor cells in monolayer and spheroid culture

Bredel-Geissler

Karbach

Walenta

et al. 1992

Journal Cellular Physiology

Self Cite

View full text Add to dashboard Cite

The oxygen consumption rate, proliferative activity, and morphology of EMT6/Ro mouse mammary sarcoma cells in monolayer and multicellular spheroid culture have been investigated in a comparative study. During the transition of monolayer cells from the exponential into the plateau growth phase, there is a distinct decrease in the cellular volume that is associated with a corresponding decrease in the proliferative and respiratory activity of the cells. The decline in cell volume is mainly due to a decrease in the content of cytoplasm, whereas the size of the nucleus is only slightly reduced. A concomitant decrease in the number of mitochondria per cell obviously accounts for the reduction in cellular oxygen uptake. Despite a continuous decrease of cell proliferation from the surface to interior regions of EMT6 spheroids reflected by a gradient in tritiated thymidine labeling, volume-related oxygen consumption is rather uniform in viable regions of these aggregates. The finding can be explained by the results of the morphometric evaluation showing a uniform volume density of mitochondria, i.e., of oxygen-consuming sites within these spheroids.

show abstract

Biological response of multicellular emt6 spheroids to exogenous lactate

Cited by 40 publications

References 26 publications

Energy metabolism transition in multi‐cellular human tumor spheroids

Energy metabolism transition in multi‐cellular human tumor spheroids

Variable presence of hypoxia in M006 human glioma spheroids and in spheroids and xenografts of clonally derived sublines

Proliferation‐associated oxygen consumption and morphology of tumor cells in monolayer and spheroid culture

Contact Info

Product

Resources

About