Methods to Measure the Inhibition of ABCG2 Transporter and Ferrochelatase Activity to Enhance Aminolevulinic Acid-Protoporphyrin IX Fluorescence-Guided Tumor Detection and Resection

Mansi, Matthew; Howley, Richard; Chen, Bin

doi:10.1007/978-1-0716-1811-0_43

Cited by 3 publications

(3 citation statements)

References 31 publications

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“…Spectrofluorometric measurement of PpIX. PpIX was measured with spectrofluorometry as described previously (22). Cells were implanted in 6-well culture plates in complete medium and allowed to attach and grow for overnight.…”

Section: Methodsmentioning

confidence: 99%

“…FECH activity assay. FECH activity was determined as described (22). Cells were implanted in 100-mm cell culture dishes and allowed to grow to about 70% confluency.…”

Section: Methodsmentioning

confidence: 99%

“…PpIX fluorescence was measured with flow cytometry as described (22). Cells were implanted in 60‐mm dishes and allowed to grow to 70% confluency.…”

Section: Methodsmentioning

confidence: 99%

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Analysis of Renal Cell Carcinoma Cell Response to the Enhancement of 5‐aminolevulinic Acid‐mediated Protoporphyrin IX Fluorescence by Iron Chelator Deferoxamine^†

Howley

Mansi

Shinde

et al. 2022

Photochem & Photobiology

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As a tumor photodiagnostic agent, 5-aminolevulinic acid (ALA) is metabolized in the heme biosynthesis pathway to produce protoporphyrin IX (PpIX) with fluorescence. ALA-PpIX fluorescence was evaluated in human renal cell carcinoma (RCC) cell lines and non-tumor HK-2 cell lines. We found that extracellular PpIX level was correlated with ABCG2 activity, illustrating its importance as a PpIX efflux transporter. Extracellular PpIX was also related to the K m of ferrochelatase (FECH) that chelates PpIX with ferrous iron to form heme. The V max of FECH was higher in all RCC cell lines tested than in the HK-2 cell line. TCGA dataset analysis indicates a positive correlation between FECH expression and RCC patient survival. These findings suggest FECH as an important biomarker in RCC. Effects of iron chelator deferoxamine (DFO) on the enhancement of PpIX fluorescence were assessed. DFO increased intracellular PpIX in both tumor and non-tumor cells, resulting in no gain in tumor/non-tumor fluorescence ratios. DFO appeared to increase ALA-PpIX more at 1-h than at 4-h treatment. There was an inverse correlation between ALA-PpIX fluorescence and the enhancement effect of DFO. These results suggest that enhancement of ALA-PpIX by DFO may be limited by the availability of ferrous iron in mitochondria following ALA administration.

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Section: Methodsmentioning

confidence: 99%

“…FECH activity assay. FECH activity was determined as described (22). Cells were implanted in 100-mm cell culture dishes and allowed to grow to about 70% confluency.…”

Section: Methodsmentioning

confidence: 99%

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Analysis of Renal Cell Carcinoma Cell Response to the Enhancement of 5‐aminolevulinic Acid‐mediated Protoporphyrin IX Fluorescence by Iron Chelator Deferoxamine^†

Howley

Mansi

Shinde

et al. 2022

Photochem & Photobiology

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Enhancing 5-ALA-PDT efficacy against resistant tumor cells: Strategies and advances

Ebrahimi,

Khaleghi Ghadiri,

Stummer

et al. 2024

Life Sciences

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The Circadian Rhythm of Intracellular Protoporphyrin IX Accumulation Through Heme Synthesis Pathway in Bladder Urothelial Cancer Cells Exposed to 5-Aminolevulinic Acid

Nishimura,

Miyake,

Onishi

et al. 2024

Cancers

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Background/Objectives: The standard recommendation for patients with non-muscle invasive bladder cancer is 5-aminolevulinic acid-mediated photodynamic diagnosis. The intensity of the fluorescence caused by the intracellular accumulation of protoporphyrin IX (PPIX) varies among tumors and patients. This study investigated the circadian rhythm of intracellular PPIX accumulation in bladder urothelial cancer cells exposed to 5-aminolevulinic acid. Methods: The expression of two clock genes, PER2 and BMAL1, and their impact on intracellular PPIX accumulation were evaluated in two bladder cancer cell lines, UM-UC-3 and J82, and mouse xenograft models. We evaluated the enzymes involved in the heme synthesis pathway that potentially affect the circadian rhythm of intracellular PPIX accumulation. The red fluorescence intensity of the images captured during photodynamic diagnosis-assisted transurethral resection of bladder tumors was quantified and compared among the four groups according to surgery start time: 9 a.m.–11 a.m., 11 a.m.–1 p.m., 1–3 p.m., and 3–5 p.m. Results: We observed the circadian rhythm of intracellular PPIX accumulation, which was potentially regulated by the clock genes PER2 and BMAL1. Two enzymes involved in the heme synthesis pathway, coproporphyrinogen oxidase and ferrochelatase, exhibit a circadian rhythm. The fluorescence intensity started gradually increasing at 12 p.m., and the highest level was observed in patients who underwent surgery between 3 and 5 p.m. Conclusions: Our findings suggest that it may be possible to optimize the timing of the photodynamic diagnosis in photodynamic diagnosis-assisted transurethral resection of bladder cancer based on the circadian rhythm to improve tumor detection and treatment outcomes.

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Methods to Measure the Inhibition of ABCG2 Transporter and Ferrochelatase Activity to Enhance Aminolevulinic Acid-Protoporphyrin IX Fluorescence-Guided Tumor Detection and Resection

Cited by 3 publications

References 31 publications

Analysis of Renal Cell Carcinoma Cell Response to the Enhancement of 5‐aminolevulinic Acid‐mediated Protoporphyrin IX Fluorescence by Iron Chelator Deferoxamine^†

Analysis of Renal Cell Carcinoma Cell Response to the Enhancement of 5‐aminolevulinic Acid‐mediated Protoporphyrin IX Fluorescence by Iron Chelator Deferoxamine^†

Enhancing 5-ALA-PDT efficacy against resistant tumor cells: Strategies and advances

The Circadian Rhythm of Intracellular Protoporphyrin IX Accumulation Through Heme Synthesis Pathway in Bladder Urothelial Cancer Cells Exposed to 5-Aminolevulinic Acid

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Methods to Measure the Inhibition of ABCG2 Transporter and Ferrochelatase Activity to Enhance Aminolevulinic Acid-Protoporphyrin IX Fluorescence-Guided Tumor Detection and Resection

Cited by 3 publications

References 31 publications

Analysis of Renal Cell Carcinoma Cell Response to the Enhancement of 5‐aminolevulinic Acid‐mediated Protoporphyrin IX Fluorescence by Iron Chelator Deferoxamine†

Analysis of Renal Cell Carcinoma Cell Response to the Enhancement of 5‐aminolevulinic Acid‐mediated Protoporphyrin IX Fluorescence by Iron Chelator Deferoxamine†

Enhancing 5-ALA-PDT efficacy against resistant tumor cells: Strategies and advances

The Circadian Rhythm of Intracellular Protoporphyrin IX Accumulation Through Heme Synthesis Pathway in Bladder Urothelial Cancer Cells Exposed to 5-Aminolevulinic Acid

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Analysis of Renal Cell Carcinoma Cell Response to the Enhancement of 5‐aminolevulinic Acid‐mediated Protoporphyrin IX Fluorescence by Iron Chelator Deferoxamine^†

Analysis of Renal Cell Carcinoma Cell Response to the Enhancement of 5‐aminolevulinic Acid‐mediated Protoporphyrin IX Fluorescence by Iron Chelator Deferoxamine^†