Protoporphyrin IX accumulation in cells treated with 5-aminolevulinic acid: Dependence on cell density, cell size and cell cycle

Moan, Johan Emelian; Bech, Øystein; Gaullier, Jean Michel; Stokke, Trond; Steen, Harald B.; Wei, Li; Berg, Kristian

doi:10.1002/(sici)1097-0215(19980105)75:1<134::aid-ijc20>3.0.co;2-f

Cited by 65 publications

(50 citation statements)

References 26 publications

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“…Specifically, we find that the highdensity populations produce substantially higher amounts of PpIX per cell than the low-density population. These results agree with fluorescence microscopy and flow cytometry studies performed by Moan et al (1998) with human colon adenocarcinoma cells and Chinese hamster lung fibroblasts. Steinbach et al (1995) observed an increasing amount of PpIX fluorescence with increasing cell density for two malignant and a normal urothelial cell line.…”

Section: Discussionsupporting

confidence: 90%

“…These results are in agreement with studies performed by Fukuda et al (1993) showing that there is little obvious cell cycle dependence on PpIX generation by CNCM-I-221 mammalian epithelial cells. Moan et al (1998) have reported that the only cell cycle dependence of PpIX synthesis appears to be a simple volume dependence, with cells in the G2+M phases producing 1.9 times more PpIX than cells in the G1 phase. The higher mean PpIX fluorescence for the fed-plateau population could be explained by cell size-dependent PpIX production; however, our experiments suggest that the differences in the number of cells corresponding to each cell cycle phase for the high-density fed-plateau and unfed-plateau cells are not significant enough to result in statistically different PpIX fluorescence measurements.…”

Section: Discussionmentioning

confidence: 99%

“…Such studies have explored the role of ALA incubation time (Dietel et al, 1996;Krammer and Uberriegler, 1996;Gibson et al, 1997) and concentration (Krammer and Uberriegler, 1996;Gibson et al, 1997;Moan et al, 1998), cell type (Iinuma et al, 1994;Steinback et al, 1995;Wyld et al, 1997), cell cycle (Moan et al, 1998), proliferation status (Rebeiz et al, 1992;Schick et al, 1995;Momma et al, 1997), serum presence (Hanania and Malik, 1992;Fukuda et al, 1993), iron availability (Hanania and Malik, 1992;Iinuma et al, 1994;Rittenhouse-Diakun et al, 1995;Tan et al, 1997), pH (Bermudez Moretti et al, 1993;Krammer and Uberriegler, 1996;Bech et al, 1997;Wyld et al, 1998), glucose (Dietel et al, 1996), and temperature (Dietel et al, 1996). More detailed studies examined the effect of some of these factors on specific enzymes of the haem biosynthetic pathway (Schoenfeld et al, 1988;Gibson et al, 1998).…”

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confidence: 99%

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Hypoxia significantly reduces aminolaevulinic acid-induced protoporphyrin IX synthesis in EMT6 cells

1999

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SummaryWe have studied the effects of hypoxia on aminolaevulinic acid (ALA)-induced protoporphyrin IX (PpIX) synthesis in EMT6 monolayer cultures characterized by different cell densities and proliferation rates. Specifically, after ALA incubation under hypoxic or normoxic conditions, we detected spectrofluorometrically the PpIX content of the following populations: (a) low-density exponentially growing cells; (b) high-density fed-plateau cells; and (c) high-density unfed-plateau cells. These populations were selected either for the purpose of comparison with other in vitro studies (low-density exponentially growing cells) or as representatives of tumour regions adjacent to (highdensity fed-plateau cells) and further away from (high-density unfed-plateau cells) capillaries. The amount of PpIX per cell produced by each one of these populations was higher after normoxic ALA incubation. The magnitude of the effect of hypoxia on PpIX synthesis was dependent on cell density and proliferation rate. A 42-fold decrease in PpIX fluorescence was observed for the high-density unfed-plateau cells. PpIX production by the low-density exponential cells was affected the least by ALA incubation under hypoxic conditions (1.4-fold decrease), whereas the effect on the high-density fed-plateau population was intermediate (20-fold decrease).

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Section: Discussionsupporting

confidence: 90%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Hypoxia significantly reduces aminolaevulinic acid-induced protoporphyrin IX synthesis in EMT6 cells

1999

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“…One difference is that they used confluent cultures exposed to δ-ALA and light while we performed experiments with cells in log phase growth. According to earlier reports (Washbrook et al, 1997;Wyld et al, 1997;Moan et al, 1998) and our unpublished results, δ-ALA uptake, PPIX accumulation and PBGD activity are dependent on cell type and cell density. This latter phenomenon could be a major confounding factor in comparisons of data obtained by different groups.…”

Section: Discussionsupporting

confidence: 70%

δ-Aminolaevulinic acid-induced photodynamic therapy inhibits protoporphyrin IX biosynthesis and reduces subsequent treatment efficacy in vitro

Havens

Nguyen

et al. 1999

Br J Cancer

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Summary Recently, considerable interest has been given to photodynamic therapy of cancer using δ-aminolaevulinic acid to induce protoporphyrin IX as the cell photosensitizer. One advantage of this modality is that protoporphyrin IX is cleared from tissue within 24 h after δ-aminolaevulinic acid administration. This could allow for multiple treatment regimens because of little concern regarding the accumulation of the photosensitizer in normal tissues. However, the haem biosynthetic pathway would have to be fully functional after the first course of therapy to allow for subsequent treatments. Photosensitization of cultured R3230AC rat mammary adenocarcinoma cells with δ-aminolaevulinic acid-induced protoporphyrin IX resulted in the inhibition of porphobilinogen deaminase, an enzyme in the haem biosynthetic pathway, and a concomitant decrease in protoporphyrin IX levels. Cultured R3230AC cells exposed to 0.5 mM δ-aminolaevulinic acid for 27 h accumulated 6.07 × 10 -16 mol of protoporphyrin IX per cell and had a porphobilinogen deaminase activity of 0.046 fmol uroporphyrin per 30 min per cell. Cells cultured under the same incubation conditions but exposed to 30 mJ cm -2 irradiation after a 3-h incubation with δ-aminolaevulinic acid showed a significant reduction in protoporphyrin IX, 2.28 × 10 -16 mol per cell, and an 80% reduction in porphobilinogen deaminase activity to 0.0088 fmol uroporphyrin per 30 min per cell. Similar effects were evident in irradiated cells incubated with δ-aminolaevulinic acid immediately after, or following a 24 h interval, post-irradiation. There was little gain in efficacy from a second treatment regimen applied within 24 h of the initial treatment, probably a result of initial metabolic damage leading to reduced levels of protoporphyrin IX. These findings suggest that a correlation may exist between the δ-aminolaevulinic acid induction of porphobilinogen deaminase activity and the increase in intracellular protoporphyrin IX accumulation.

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“…Cell density is known to affect the rate of PpIX generation. Increasing tumour cell density by a factor of 10 increases PpIX production (Moan et al, 1998). The present results show a reverse trend in PpIX production despite the higher density of cells in the high glucose cells (30610 5 cells ml 71 at 0 mM glucose vs 44610 5 cells ml 71 at 25 mM glucose), suggesting the involvement of additional processes.…”

Section: Discussioncontrasting

confidence: 47%

Aminolaevulinic acid-induced photodynamic therapy: cellular responses to glucose starvation

et al. 2002

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Photodynamic therapy is a cancer treatment based on the interaction of light, oxygen and a photosensitiser. Protoporphyrin. IX is an endogenous photosensitiser derived from the pro-drug aminolaevulinic acid. Tumours contain areas of hypoxia and hypoglycaemia. Tumour cells adapt to these conditions by stress protein induction which may induce resistance to cancer therapies. The effect of chronic hypoglycaemia on sensitivity to aminolaevulinic acid-induced photodynamic therapy in vitro was studied in MCF-7, human breast cancer cells. Following chronic exposure to 0, 1 or 25 mM, glucose, cells were treated with aminolaevulinic acid and the generation of intracellular protoporphyrin. IX measured by spectrofluorimetry. Aminolaevulinic acid-induced photodynamic therapy sensitivity was compared between cells following chronic exposure to 0, 1 or 25 mM glucose. Percentage cell survival was determined by clonogenic assay. Cells cultured in low glucose generated higher levels of protoporphyrin IX compared to standard glucose medium (0 mM glucose: 0.88610 75 ng cell 71 , 1 mM: 0.86610 75 ng cell 71 , 25 mM: 0.605610 75 ng cell 71 , P50.05). However, photodynamic therapy sensitivity was reduced in glucose deprived cells (0 mM glucose: 61% survival, 1 mM: 80.5% and 25 mM: 39.6%, P50.05). Chronic exposure to low glucose induces photodynamic therapy resistance despite increased intracellular concentrations of protoporphyrin IX and may reflect cellular adaptation to chronic glucose deprivation.

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Protoporphyrin IX accumulation in cells treated with 5-aminolevulinic acid: Dependence on cell density, cell size and cell cycle

Cited by 65 publications

References 26 publications

Hypoxia significantly reduces aminolaevulinic acid-induced protoporphyrin IX synthesis in EMT6 cells

Hypoxia significantly reduces aminolaevulinic acid-induced protoporphyrin IX synthesis in EMT6 cells

δ-Aminolaevulinic acid-induced photodynamic therapy inhibits protoporphyrin IX biosynthesis and reduces subsequent treatment efficacy in vitro

Aminolaevulinic acid-induced photodynamic therapy: cellular responses to glucose starvation

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