Methotrexate enhances 5-aminolevulinic acid-mediated photodynamic therapy-induced killing of human SCC4 cells by upregulation of coproporphyrinogen oxidase
Topical 5-aminolevulinic acid-mediated photodynamic therapy (ALA-PDT) is effective for treatment of oral precancerous and cancerous lesions. This in vitro study tried to examine whether the SCC4 cell killing by ALA-PDT was enhanced by pretreatment of methotrexate (MTX). Methods: To measure the SCC4...
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| Format: | Article |
| Language: | English |
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Elsevier
2014-02-01
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| Series: | Journal of the Formosan Medical Association |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S092966461300452X |
| _version_ | 1819062136801853440 |
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| author | Deng-Fu Yang Jeng-Woei Lee Hsin-Ming Chen Zheng Huang Yih-Chih Hsu |
| author_facet | Deng-Fu Yang Jeng-Woei Lee Hsin-Ming Chen Zheng Huang Yih-Chih Hsu |
| author_sort | Deng-Fu Yang |
| collection | DOAJ |
| description | Topical 5-aminolevulinic acid-mediated photodynamic therapy (ALA-PDT) is effective for treatment of oral precancerous and cancerous lesions. This in vitro study tried to examine whether the SCC4 cell killing by ALA-PDT was enhanced by pretreatment of methotrexate (MTX).
Methods: To measure the SCC4 cell killing abilities by MTX-pretreated ALA-PDT (MTX–ALA-PDT), the SCC4 cells were pretreated with 0 mg/L, 0.001 mg/L, 0.01 mg/L, 0.1 mg/L, or 1 mg/L of MTX for 72 hours, then incubated with 0 mM, 0.0625 mM, 0.125 mM, 0.187 mM, 0.25 mM, or 0.375 mM ALA for 4 hours, and subsequently illuminated with a 640-nm light-emitting diode array at a light dose of 10 J/cm2. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was conducted at 24 hours to quantify SCC4 cell survival rates after MTX–ALA-PDT treatment. Western blot analyses were used to examine the MTX-mediated enhancement in the expressions of the heme production-related enzymes, coproporphyrinogen oxidase (CPOX), protoporphyrinogen oxidase (PPOX), and ferrochelatase, in the MTX-preconditioned SCC4 cells.
Results: Pretreatment of SCC4 cells by 0.001 mg/L MTX for 72 hours resulted in a significant augmentation in MTX–ALA-PDT-induced killing of SCC4 cells (p < 0.05). The SCC4 cells treated with 0.001 mg/L MTX for 72 hours showed a significant and 1.65-fold increase in CPOX expression compared with the control SCC4 cells without MTX treatment (p < 0.05). However, no significant changes in the expressions of PPOX and ferrochelatase were observed in the SCC4 cells pretreated with different concentrations of MTX.
Conclusion: MTX enhances ALA-PDT-induced SCC4 cell killing through upregulation of CPOX expression and subsequent increase in intracellular protoporphyrin IX production in SCC4 cells. |
| first_indexed | 2024-12-21T14:53:59Z |
| format | Article |
| id | doaj.art-89f4a53d7ec344bebcf5d4846bdd898a |
| institution | Directory Open Access Journal |
| issn | 0929-6646 |
| language | English |
| last_indexed | 2024-12-21T14:53:59Z |
| publishDate | 2014-02-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of the Formosan Medical Association |
| spelling | doaj.art-89f4a53d7ec344bebcf5d4846bdd898a2022-12-21T18:59:48ZengElsevierJournal of the Formosan Medical Association0929-66462014-02-011132889310.1016/j.jfma.2013.12.005Methotrexate enhances 5-aminolevulinic acid-mediated photodynamic therapy-induced killing of human SCC4 cells by upregulation of coproporphyrinogen oxidaseDeng-Fu Yang0Jeng-Woei Lee1Hsin-Ming Chen2Zheng Huang3Yih-Chih Hsu4Graduate Program, Department of Bioscience Technology, Chung Yuan Christian University, Taoyuan, TaiwanDepartment of Life Science, Tzu Chi University, Hualien, TaiwanGraduate Institute of Oral Biology, School of Dentistry, National Taiwan University, Taipei, TaiwanCancer Center, University of Colorado Denver Cancer Center, CO, USADepartment of Bioscience Technology, Chung Yuan Christian University, Taoyuan, TaiwanTopical 5-aminolevulinic acid-mediated photodynamic therapy (ALA-PDT) is effective for treatment of oral precancerous and cancerous lesions. This in vitro study tried to examine whether the SCC4 cell killing by ALA-PDT was enhanced by pretreatment of methotrexate (MTX). Methods: To measure the SCC4 cell killing abilities by MTX-pretreated ALA-PDT (MTX–ALA-PDT), the SCC4 cells were pretreated with 0 mg/L, 0.001 mg/L, 0.01 mg/L, 0.1 mg/L, or 1 mg/L of MTX for 72 hours, then incubated with 0 mM, 0.0625 mM, 0.125 mM, 0.187 mM, 0.25 mM, or 0.375 mM ALA for 4 hours, and subsequently illuminated with a 640-nm light-emitting diode array at a light dose of 10 J/cm2. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was conducted at 24 hours to quantify SCC4 cell survival rates after MTX–ALA-PDT treatment. Western blot analyses were used to examine the MTX-mediated enhancement in the expressions of the heme production-related enzymes, coproporphyrinogen oxidase (CPOX), protoporphyrinogen oxidase (PPOX), and ferrochelatase, in the MTX-preconditioned SCC4 cells. Results: Pretreatment of SCC4 cells by 0.001 mg/L MTX for 72 hours resulted in a significant augmentation in MTX–ALA-PDT-induced killing of SCC4 cells (p < 0.05). The SCC4 cells treated with 0.001 mg/L MTX for 72 hours showed a significant and 1.65-fold increase in CPOX expression compared with the control SCC4 cells without MTX treatment (p < 0.05). However, no significant changes in the expressions of PPOX and ferrochelatase were observed in the SCC4 cells pretreated with different concentrations of MTX. Conclusion: MTX enhances ALA-PDT-induced SCC4 cell killing through upregulation of CPOX expression and subsequent increase in intracellular protoporphyrin IX production in SCC4 cells.http://www.sciencedirect.com/science/article/pii/S092966461300452X5-aminolevulinic acidcoproporphyrinogen oxidaseferrochelatasehuman squamous cell carcinoma 4 cellmethotrexatephotodynamic therapy |
| spellingShingle | Deng-Fu Yang Jeng-Woei Lee Hsin-Ming Chen Zheng Huang Yih-Chih Hsu Methotrexate enhances 5-aminolevulinic acid-mediated photodynamic therapy-induced killing of human SCC4 cells by upregulation of coproporphyrinogen oxidase Journal of the Formosan Medical Association 5-aminolevulinic acid coproporphyrinogen oxidase ferrochelatase human squamous cell carcinoma 4 cell methotrexate photodynamic therapy |
| title | Methotrexate enhances 5-aminolevulinic acid-mediated photodynamic therapy-induced killing of human SCC4 cells by upregulation of coproporphyrinogen oxidase |
| title_full | Methotrexate enhances 5-aminolevulinic acid-mediated photodynamic therapy-induced killing of human SCC4 cells by upregulation of coproporphyrinogen oxidase |
| title_fullStr | Methotrexate enhances 5-aminolevulinic acid-mediated photodynamic therapy-induced killing of human SCC4 cells by upregulation of coproporphyrinogen oxidase |
| title_full_unstemmed | Methotrexate enhances 5-aminolevulinic acid-mediated photodynamic therapy-induced killing of human SCC4 cells by upregulation of coproporphyrinogen oxidase |
| title_short | Methotrexate enhances 5-aminolevulinic acid-mediated photodynamic therapy-induced killing of human SCC4 cells by upregulation of coproporphyrinogen oxidase |
| title_sort | methotrexate enhances 5 aminolevulinic acid mediated photodynamic therapy induced killing of human scc4 cells by upregulation of coproporphyrinogen oxidase |
| topic | 5-aminolevulinic acid coproporphyrinogen oxidase ferrochelatase human squamous cell carcinoma 4 cell methotrexate photodynamic therapy |
| url | http://www.sciencedirect.com/science/article/pii/S092966461300452X |
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