Skip to main content
Top
Published in: Lasers in Medical Science 1/2011

01-01-2011 | Original Article

Effect and mechanism of 5-aminolevulinic acid-mediated photodynamic therapy in esophageal cancer

Authors: Xiaohua Chen, Peng Zhao, Fengsheng Chen, Libo Li, Rongcheng Luo

Published in: Lasers in Medical Science | Issue 1/2011

Login to get access

Abstract

5-aminolevulinic acid-mediated photodynamic therapy (ALA-PDT) provides a novel and promising treatment for esophageal cancer. However, its specific mechanism has not been fully elucidated and its efficacy is remarkably varied. This study investigated the effect of ALA-PDT on esophageal squamous carcinoma cell line Eca-109 in vitro and vivo to explore optimal parameters, and evaluated the significance of cell apoptosis, cell cycle, ALA-protoporphyrin IX (ALA-PpIX) subcellular localization, and expression of Bcl-2 and Bax mRNA in cells to understand the mechanism of ALA-PDT for esophageal cancer. How ALA concentration, incubation time, and laser irradiation dose influenced the cell proliferation was determined by MTT assay. ALA-PpIX subcellular localization was analyzed by confocal microscopy. The mRNA changes were detected by quantitative real-time polymerase chain reaction (QRT-PCR). Tumor models transplanted with Eca-109 cells in nude mice were established (n = 10) and killed (n = 4) at 24 h post-PDT for malondialdehyde (MDA) detection and histological study. The remaining mice were measured the tumor size for 3 weeks after treatment. Our data show that ALA-PDT significantly inhibits cell proliferation (p < 0.05), the PDT efficacy depends on the saturation of ALA concentration, incubation time, and laser irradiation dose, and the best effect in tumor destruction is at 7–14 days post-PDT. ALA-PpIX is localized in mitochondria and cytoplasm. ALA-PDT induces cell apoptosis and arrests cell cycle at G0/G1 phase. Bcl-2 is significantly down-regulated while Bax is up-regulated (p < 0.05). The results of this study provide references in choosing clinical optimal PDT parameters and help in better understanding the PDT mechanism for esophageal cancer.
Literature
1.
go back to reference Pisani P, Parkin DM, Bray F, Ferlay J (1999) Estimates of the worldwide mortality from 25 cancers in 1990. Int J Cancer 83:18–29CrossRefPubMed Pisani P, Parkin DM, Bray F, Ferlay J (1999) Estimates of the worldwide mortality from 25 cancers in 1990. Int J Cancer 83:18–29CrossRefPubMed
2.
go back to reference Tran GD, Sun XD, Abnet CC, Fan JH, Dawsey SM, Dong ZW, Mark SD, Qiao YL, Taylor PR (2005) Prospective study of risk factors for esophageal and gastric cancers in the Linxian general population trial cohort in China. Int J Cancer 113:456–463. doi:10.1002/ijc.20616 CrossRefPubMed Tran GD, Sun XD, Abnet CC, Fan JH, Dawsey SM, Dong ZW, Mark SD, Qiao YL, Taylor PR (2005) Prospective study of risk factors for esophageal and gastric cancers in the Linxian general population trial cohort in China. Int J Cancer 113:456–463. doi:10.​1002/​ijc.​20616 CrossRefPubMed
3.
go back to reference Litle VR, Luketich JD, Christie NA, Buenaventura PO, Alvelo-Rivera M, McCaughan JS, Nguyen NT, Fernando HC (2003) Photodynamic therapy as palliation for esophageal cancer: experience in 215 patients. Ann Thorac Surg 76:1687–1693. doi:10.1016/S0003-4975(03)01299-2 CrossRefPubMed Litle VR, Luketich JD, Christie NA, Buenaventura PO, Alvelo-Rivera M, McCaughan JS, Nguyen NT, Fernando HC (2003) Photodynamic therapy as palliation for esophageal cancer: experience in 215 patients. Ann Thorac Surg 76:1687–1693. doi:10.​1016/​S0003-4975(03)01299-2 CrossRefPubMed
4.
go back to reference Moghissi K, Dixon K, Thorpe JAC, Stringer M, Moore PJ (2000) The role of photodynamic therapy in inoperable oesophageal cancer. Eur J Cardiothorac Surg 17:95–100CrossRefPubMed Moghissi K, Dixon K, Thorpe JAC, Stringer M, Moore PJ (2000) The role of photodynamic therapy in inoperable oesophageal cancer. Eur J Cardiothorac Surg 17:95–100CrossRefPubMed
6.
go back to reference Manyak MJ, Russo A, Smith PD, Glatstein E (1998) Photodynamic therapy. J Clin Oncol 6:380–391 Manyak MJ, Russo A, Smith PD, Glatstein E (1998) Photodynamic therapy. J Clin Oncol 6:380–391
7.
go back to reference McCaughan JS Jr, Ellison EC, Guy JT, Hicks WJ, MD JJJ, Laufman LR, May E, Nims TA, Spiridonidis CH, Williams TE (1996) Photodynamic therapy for esophageal malignancy: a prospective twelve-year study. Ann Thorac Surg 62:1005–1010CrossRefPubMed McCaughan JS Jr, Ellison EC, Guy JT, Hicks WJ, MD JJJ, Laufman LR, May E, Nims TA, Spiridonidis CH, Williams TE (1996) Photodynamic therapy for esophageal malignancy: a prospective twelve-year study. Ann Thorac Surg 62:1005–1010CrossRefPubMed
8.
go back to reference Hopper C (2000) Photodynamic therapy: a clinical reality in the treatment of cancer. Lancet Oncol 1:212–219CrossRefPubMed Hopper C (2000) Photodynamic therapy: a clinical reality in the treatment of cancer. Lancet Oncol 1:212–219CrossRefPubMed
9.
go back to reference Dougherty TJ, Gomer CJ, Henderson BW, Jori G, Kessel D, Korbelik M, Moan J, Peng Q (1998) Photodynamic therapy. J Natl Cancer Inst 90:889–905CrossRefPubMed Dougherty TJ, Gomer CJ, Henderson BW, Jori G, Kessel D, Korbelik M, Moan J, Peng Q (1998) Photodynamic therapy. J Natl Cancer Inst 90:889–905CrossRefPubMed
11.
go back to reference Luo Y, Kessel D (1997) Initiation of apoptosis versus necrosis by photodynamic therapy with chloroaluminum phthalocyanine. Photochem Photobiol 66:479–483CrossRefPubMed Luo Y, Kessel D (1997) Initiation of apoptosis versus necrosis by photodynamic therapy with chloroaluminum phthalocyanine. Photochem Photobiol 66:479–483CrossRefPubMed
12.
go back to reference Lavie G, Kaplinsky C, Toren A, Aizman I, Meruelo D, Mazur Y, Mandel M (1999) A photodynamic pathway to apoptosis and necrosis induced by dimethyl tetrahydroxyhelianthrone and hypericin in leukaemic cells: possible relevance to photodynamic therapy. Br J Cancer 79:423–432CrossRefPubMed Lavie G, Kaplinsky C, Toren A, Aizman I, Meruelo D, Mazur Y, Mandel M (1999) A photodynamic pathway to apoptosis and necrosis induced by dimethyl tetrahydroxyhelianthrone and hypericin in leukaemic cells: possible relevance to photodynamic therapy. Br J Cancer 79:423–432CrossRefPubMed
14.
go back to reference Ali SM, Olivo M (2002) Bio-distribution and subcellular localization of Hypericin and its role in PDT induced apoptosis in cancer cells. Int J Oncol 21:531–540PubMed Ali SM, Olivo M (2002) Bio-distribution and subcellular localization of Hypericin and its role in PDT induced apoptosis in cancer cells. Int J Oncol 21:531–540PubMed
15.
go back to reference Murant RS, Gibson SL, Hilf R (1987) Photosensitizing effects of Photofrin II on the site selected mitochondrial enzymes adenylate kinase and monoamine oxidase. Cancer Res 47:4323–4328PubMed Murant RS, Gibson SL, Hilf R (1987) Photosensitizing effects of Photofrin II on the site selected mitochondrial enzymes adenylate kinase and monoamine oxidase. Cancer Res 47:4323–4328PubMed
16.
go back to reference Woodburn KW, Vardaxis NJ, Hill JS, Kaye AH, Phillips DR (1991) Subcellular localization of porphyrins using confocal laser scanning microscopy. Photochem Photobiol 54:725–732CrossRefPubMed Woodburn KW, Vardaxis NJ, Hill JS, Kaye AH, Phillips DR (1991) Subcellular localization of porphyrins using confocal laser scanning microscopy. Photochem Photobiol 54:725–732CrossRefPubMed
17.
go back to reference Ricchelli F, Gobbo S, Jori G, Salet C, Moreno G (1995) Temperature induced changes in fluorescence properties as a probe of porphyrin microenvironment in lipid membranes: 2. The partition of hematoporphyrin and protoporphyrin in mitochondria. Eur J Biochem 233:165–170CrossRefPubMed Ricchelli F, Gobbo S, Jori G, Salet C, Moreno G (1995) Temperature induced changes in fluorescence properties as a probe of porphyrin microenvironment in lipid membranes: 2. The partition of hematoporphyrin and protoporphyrin in mitochondria. Eur J Biochem 233:165–170CrossRefPubMed
18.
go back to reference Szeimies RM, Karrer S, Abels C, Steinbach P, Fickweiler S, Messmann H, Baumler W, Landthaler M (1996) 9-Acetoxy-2, 7, 12, 17- tetrakis-(beta-methoxyethyl)-porphycene (ATMPn), a novel photosensitizer for photodynamic therapy: uptake kinetics and intracellular localization. J Photochem Photobiol B 34:67–72CrossRefPubMed Szeimies RM, Karrer S, Abels C, Steinbach P, Fickweiler S, Messmann H, Baumler W, Landthaler M (1996) 9-Acetoxy-2, 7, 12, 17- tetrakis-(beta-methoxyethyl)-porphycene (ATMPn), a novel photosensitizer for photodynamic therapy: uptake kinetics and intracellular localization. J Photochem Photobiol B 34:67–72CrossRefPubMed
19.
go back to reference Graham A, Li G, Chen Y, Morgan J, Oseroff A, Dougherty TJ, Pandey RK (2003) Structure–activity relationship of new octaethylporphyrin-based benzochlorins as photosensitizers for photodynamic therapy. Photochem Photobiol 77:561–566PubMed Graham A, Li G, Chen Y, Morgan J, Oseroff A, Dougherty TJ, Pandey RK (2003) Structure–activity relationship of new octaethylporphyrin-based benzochlorins as photosensitizers for photodynamic therapy. Photochem Photobiol 77:561–566PubMed
20.
go back to reference Tsai JC, Wu CL, Chien HF, Chen CT (2005) Reorganization of cytoskeleton induced by 5-aminolevulinic acid-mediated photodynamic therapy and its correlation with mitochondrial dysfunction. Lasers Surg Med 36:398–408. doi:10.1002/lsm.20179 CrossRefPubMed Tsai JC, Wu CL, Chien HF, Chen CT (2005) Reorganization of cytoskeleton induced by 5-aminolevulinic acid-mediated photodynamic therapy and its correlation with mitochondrial dysfunction. Lasers Surg Med 36:398–408. doi:10.​1002/​lsm.​20179 CrossRefPubMed
21.
go back to reference Lu Y, Jiao RQ, Chen XP, Zhong JY, Ji JG, Shen PP (2008) Methylene blue-mediated photodynamic therapy induces mitochondria-dependent apoptosis in HeLa cell. J Cell Biochem 105:1451–1460. doi:10.1002/jcb.21965 CrossRefPubMed Lu Y, Jiao RQ, Chen XP, Zhong JY, Ji JG, Shen PP (2008) Methylene blue-mediated photodynamic therapy induces mitochondria-dependent apoptosis in HeLa cell. J Cell Biochem 105:1451–1460. doi:10.​1002/​jcb.​21965 CrossRefPubMed
22.
go back to reference Lu ZB, Tao Y, Zhou ZX, Zhang JJ, Li C, Ou LC, Zhao BL (2006) Mitochondrial reactive oxygen species and nitric oxide-mediated cancer cell apoptosis in 2-butylamino-2-demethoxyhypocrellin B photodynamic treatment. Free Radic Biol Med 41:1590–1605CrossRefPubMed Lu ZB, Tao Y, Zhou ZX, Zhang JJ, Li C, Ou LC, Zhao BL (2006) Mitochondrial reactive oxygen species and nitric oxide-mediated cancer cell apoptosis in 2-butylamino-2-demethoxyhypocrellin B photodynamic treatment. Free Radic Biol Med 41:1590–1605CrossRefPubMed
23.
go back to reference Lam M, Oleinick NL, Nieminen AL (2001) Photodynamic therapy-induced apoptosis in epidermoid carcinoma cells. Reactive oxygen species and mitochondrial inner membrane permeabilization. J Biol Chem 276:47379–47386. doi:10.1074/jbc.M107678200 CrossRefPubMed Lam M, Oleinick NL, Nieminen AL (2001) Photodynamic therapy-induced apoptosis in epidermoid carcinoma cells. Reactive oxygen species and mitochondrial inner membrane permeabilization. J Biol Chem 276:47379–47386. doi:10.​1074/​jbc.​M107678200 CrossRefPubMed
24.
go back to reference Tsai T, Ji HT, Chiang PC, Chou RH, Chang WSW, Chen CT (2009) ALA-PDT results in phenotypic changes and decreased cellular invasion in surviving cancer cells. Lasers Surg Med 41:305–315. doi:10.1002/lsm.20761 CrossRefPubMed Tsai T, Ji HT, Chiang PC, Chou RH, Chang WSW, Chen CT (2009) ALA-PDT results in phenotypic changes and decreased cellular invasion in surviving cancer cells. Lasers Surg Med 41:305–315. doi:10.​1002/​lsm.​20761 CrossRefPubMed
25.
go back to reference Chen YJ, Zheng W, Li YQ, Zhong JY, Ji JG, Shen PP (2008) Apoptosis induced by methylene-blue-mediated photodynamic therapy in melanomas and the involvement of mitochondrial dysfunction revealed by proteomics. Cancer Sci 99:2019–2027. doi:10.1111/j.1349-7006.2008.00910.x PubMed Chen YJ, Zheng W, Li YQ, Zhong JY, Ji JG, Shen PP (2008) Apoptosis induced by methylene-blue-mediated photodynamic therapy in melanomas and the involvement of mitochondrial dysfunction revealed by proteomics. Cancer Sci 99:2019–2027. doi:10.​1111/​j.​1349-7006.​2008.​00910.​x PubMed
26.
go back to reference Usuda J, Chiu SM, Murphy ES, Lam M, Nieminen AL, Oleinick NL (2003) Domain dependent photodamage to Bcl-2. A membrane anchorage region is needed to form the target of phthalocyanine photosensitization. J Biol Chem 278:2021–2029. doi:10.1074/jbc.M205219200 CrossRefPubMed Usuda J, Chiu SM, Murphy ES, Lam M, Nieminen AL, Oleinick NL (2003) Domain dependent photodamage to Bcl-2. A membrane anchorage region is needed to form the target of phthalocyanine photosensitization. J Biol Chem 278:2021–2029. doi:10.​1074/​jbc.​M205219200 CrossRefPubMed
27.
go back to reference Wild PJ, Krieg RC, Seidl J, Stoehr R, Reher K, Hofmann C, Louhelainen J, Rosenthal A, Hartmann A, Pilarsky C, Bosserhoff AK, Knueche R (2005) RNA expression profiling of normal and tumor cells following photodynamic therapy with 5-aminolevulinic acid-induced protoporphyrin IX in vitro. Mol Cancer Ther 4:516–528CrossRefPubMed Wild PJ, Krieg RC, Seidl J, Stoehr R, Reher K, Hofmann C, Louhelainen J, Rosenthal A, Hartmann A, Pilarsky C, Bosserhoff AK, Knueche R (2005) RNA expression profiling of normal and tumor cells following photodynamic therapy with 5-aminolevulinic acid-induced protoporphyrin IX in vitro. Mol Cancer Ther 4:516–528CrossRefPubMed
28.
go back to reference McGarrity TJ, Peiffer LP, Granville DJ, Carthy CM, Levy JG, Khandelwal M, Hunt DWC (2001) Apoptosis associated with esophageal adenocarcinoma: influence of photodynamic therapy. Cancer Lett 163:33–41CrossRefPubMed McGarrity TJ, Peiffer LP, Granville DJ, Carthy CM, Levy JG, Khandelwal M, Hunt DWC (2001) Apoptosis associated with esophageal adenocarcinoma: influence of photodynamic therapy. Cancer Lett 163:33–41CrossRefPubMed
29.
go back to reference Matroule JY, Carthy CM, Granville DJ, Jolois O, Hunt DWC, Piette J (2001) Mechanism of colon cancer cell apoptosis mediated by pyropheophorbide-a methylester photosensitization. Oncogene 20:4070–4084CrossRefPubMed Matroule JY, Carthy CM, Granville DJ, Jolois O, Hunt DWC, Piette J (2001) Mechanism of colon cancer cell apoptosis mediated by pyropheophorbide-a methylester photosensitization. Oncogene 20:4070–4084CrossRefPubMed
30.
go back to reference Usuda J, Chiu SM, Azizuddin K, Xue LY, Lam M, Nieminen AL, Oleinick NL (2002) Promotion of photodynamic therapy-induced apoptosis by the mitochondrial protein Smac/DIABLO: dependence on Bax. Photochem Photobiol 76:217–223CrossRefPubMed Usuda J, Chiu SM, Azizuddin K, Xue LY, Lam M, Nieminen AL, Oleinick NL (2002) Promotion of photodynamic therapy-induced apoptosis by the mitochondrial protein Smac/DIABLO: dependence on Bax. Photochem Photobiol 76:217–223CrossRefPubMed
31.
go back to reference Kesse D, Reiners JJ Jr (2002) Apoptotic response to photodynamic therapy versus the Bcl-2 antagonist HA14-1. Photochem Photobiol 76:314–319CrossRef Kesse D, Reiners JJ Jr (2002) Apoptotic response to photodynamic therapy versus the Bcl-2 antagonist HA14-1. Photochem Photobiol 76:314–319CrossRef
32.
go back to reference He J, Agarwal ML, Larkin HE, Friedman LR, Xue LY, Oleinick NL (1996) The induction of partial resistance to photodynamic therapy by the protooncogene BCL-2. Photochem Photobiol 64:845–852CrossRefPubMed He J, Agarwal ML, Larkin HE, Friedman LR, Xue LY, Oleinick NL (1996) The induction of partial resistance to photodynamic therapy by the protooncogene BCL-2. Photochem Photobiol 64:845–852CrossRefPubMed
33.
go back to reference Kim HRC, Luo Y, Li G, Kessel D (1999) Enhanced apoptotic response to photodynamic therapy after bcl-2 transfection. Cancer Res 59:3429–3432PubMed Kim HRC, Luo Y, Li G, Kessel D (1999) Enhanced apoptotic response to photodynamic therapy after bcl-2 transfection. Cancer Res 59:3429–3432PubMed
34.
go back to reference Kessel D, Castelli M (2001) Evidence that bcl-2 is the target of three photosensitizers that induce a rapid apoptotic response. Photochem Photobiol 74:318–322CrossRefPubMed Kessel D, Castelli M (2001) Evidence that bcl-2 is the target of three photosensitizers that induce a rapid apoptotic response. Photochem Photobiol 74:318–322CrossRefPubMed
35.
go back to reference Au CM, Luk SK, Jackson CJ, Ng HK, Yow CMN, To SST (2006) Differential effects of photofrin, 5-aminolevulinic acid and calphostin C on glioma cells. J Photochem Photobiol B 85:92–101CrossRefPubMed Au CM, Luk SK, Jackson CJ, Ng HK, Yow CMN, To SST (2006) Differential effects of photofrin, 5-aminolevulinic acid and calphostin C on glioma cells. J Photochem Photobiol B 85:92–101CrossRefPubMed
37.
go back to reference Ruhdorfer S, Sanovic R, Sander V, Krammer B, Verwanger T (2007) Gene expression profiling of the human carcinoma cell line A-431 after 5-aminolevulinic acid-based photodynamic treatment. Int J Oncol 30:1253–1262PubMed Ruhdorfer S, Sanovic R, Sander V, Krammer B, Verwanger T (2007) Gene expression profiling of the human carcinoma cell line A-431 after 5-aminolevulinic acid-based photodynamic treatment. Int J Oncol 30:1253–1262PubMed
38.
go back to reference Casas A, Perotti C, Saccoliti M, Sacca P, Fukuda H, Batlle AMC (2002) ALA and ALA hexyl ester in free and liposomal formulations for the photosensitization of tumor organ cultures. Br J Cancer 86:837–842CrossRefPubMed Casas A, Perotti C, Saccoliti M, Sacca P, Fukuda H, Batlle AMC (2002) ALA and ALA hexyl ester in free and liposomal formulations for the photosensitization of tumor organ cultures. Br J Cancer 86:837–842CrossRefPubMed
41.
go back to reference Harris MH, Thompson CB (2000) The role of the Bcl-2 family in the regulation of outer mitochondrial membrane permeability. Cell Death Differ 7:1182–1191CrossRefPubMed Harris MH, Thompson CB (2000) The role of the Bcl-2 family in the regulation of outer mitochondrial membrane permeability. Cell Death Differ 7:1182–1191CrossRefPubMed
42.
go back to reference Kessel D, Luguya R, Vicente MGH (2003) Localization and photodynamic efficacy of two cationic porphyrins varying in charge distribution. Photochem Photobiol 78:431–435CrossRefPubMed Kessel D, Luguya R, Vicente MGH (2003) Localization and photodynamic efficacy of two cationic porphyrins varying in charge distribution. Photochem Photobiol 78:431–435CrossRefPubMed
43.
go back to reference Zhang WG, Ma LP, Wang SW, Zhang ZY, Cao GD (1999) Antisense Bcl-2 retrovirus vector increases the sensitivity of a human gastric adenocarcinoma cell line to photodynamic therapy. Photochem Photobiol 69:582–586CrossRefPubMed Zhang WG, Ma LP, Wang SW, Zhang ZY, Cao GD (1999) Antisense Bcl-2 retrovirus vector increases the sensitivity of a human gastric adenocarcinoma cell line to photodynamic therapy. Photochem Photobiol 69:582–586CrossRefPubMed
44.
go back to reference Srivastava M, Ahmad N, Gupta S, Mukhtar H (2001) Involvement of Bcl-2 and Bax in photodynamic therapy-mediated apoptosis. Antisense Bcl-2 oligonucleotide sensitizes RIF1 cells to photodynamic therapy apoptosis. J Biol Chem 276:15481–15488. doi:10.1074/jbc.M006920200 CrossRefPubMed Srivastava M, Ahmad N, Gupta S, Mukhtar H (2001) Involvement of Bcl-2 and Bax in photodynamic therapy-mediated apoptosis. Antisense Bcl-2 oligonucleotide sensitizes RIF1 cells to photodynamic therapy apoptosis. J Biol Chem 276:15481–15488. doi:10.​1074/​jbc.​M006920200 CrossRefPubMed
Metadata
Title
Effect and mechanism of 5-aminolevulinic acid-mediated photodynamic therapy in esophageal cancer
Authors
Xiaohua Chen
Peng Zhao
Fengsheng Chen
Libo Li
Rongcheng Luo
Publication date
01-01-2011
Publisher
Springer-Verlag
Published in
Lasers in Medical Science / Issue 1/2011
Print ISSN: 0268-8921
Electronic ISSN: 1435-604X
DOI
https://doi.org/10.1007/s10103-010-0810-0

Other articles of this Issue 1/2011

Lasers in Medical Science 1/2011 Go to the issue