5-Aminolevulinic acid strongly enhances delayed intracellular production of reactive oxygen species (ROS) generated by ionizing irradiation: Quantitative analyses and visualization of intracellular ROS production in glioma cells in vitro

Kitagawa,Takehiro; Yamamoto,Junkoh; Tanaka,Tohru; Nakano,Yoshiteru; Akiba,Daisuke; Ueta,Kunihiro; Nishizawa,Shigeru

doi:10.3892/or.2014.3618

5-Aminolevulinic acid strongly enhances delayed intracellular production of reactive oxygen species (ROS) generated by ionizing irradiation: Quantitative analyses and visualization of intracellular ROS production in glioma cells in vitro

Authors:
- Takehiro Kitagawa
- Junkoh Yamamoto
- Tohru Tanaka
- Yoshiteru Nakano
- Daisuke Akiba
- Kunihiro Ueta
- Shigeru Nishizawa
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Affiliations: Department of Neurosurgery, University of Occupational and Environmental Health, Kitakyushu, Fukuoka, Japan, SBI Pharmaceuticals Co., Ltd., Minato-ku, Tokyo, Japan
Published online on: November 24, 2014 https://doi.org/10.3892/or.2014.3618
Pages: 583-590

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Abstract

Postoperative adjuvant radiotherapy has important roles in multimodal treatment for highly aggressive malignant gliomas. Previously, we demonstrated that multi-dose ionizing irradiation with repetitive administration of 5-aminolevulinic acid (5-ALA) enhanced the host antitumor response and strongly inhibited tumor growth in experimental glioma. However, the mechanism of the radiosensitizing effect of 5-ALA is not known. Ionizing irradiation not only causes reactive oxygen species (ROS) formation initially by water radiolysis but also induces delayed production of mitochondrial ROS for mediating the long-lasting effects of ionizing irradiation on tumor cells. 5-ALA leads to high accumulation of protoporphyrin IX (PpIX) in the mitochondria of tumor cells, yet can also improve dysfunction of the mitochondrial respiratory chain in tumor cells. Here, we assessed the effect of 5-ALA-induced PpIX synthesis and delayed production of intracellular ROS after ionizing irradiation with 5-ALA in glioma cells in vitro. Temporal changes in intracellular 5-ALA-induced PpIX synthesis after ionizing irradiation in glioma cell lines were evaluated using flow cytometry (FCM). Then, the effect of 5-ALA on delayed production of intracellular ROS 12 h after ionizing irradiation in glioma cells was evaluated by FCM and confocal laser scanning microscopy. Ionizing irradiation had no effect on 5-ALA-induced PpIX synthesis in glioma cells. Delayed intracellular production of ROS was significantly higher than that just after ionizing irradiation, but 5-ALA pretreatment strongly enhanced the delayed intracellular production of ROS, mainly in the cytoplasm of glioma cells. This 5-ALA-induced increase in the delayed production of ROS tended to be higher in the case of 5-ALA treatment before rather than after ionizing irradiation. These results suggest that 5-ALA can affect tumor cells under ionizing irradiation, and greatly increase secondary intracellular production of ROS long after ionizing irradiation, thereby causing a radiosensitizing effect in glioma cells.

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1	Stupp R, Mason WP, van den Bent MJ, et al: Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. N Engl J Med. 352:987–996. 2005. View Article : Google Scholar : PubMed/NCBI
2	Westphal M, Ram Z, Riddle V, Hilt D and Bortey E; Executive Committee of the Gliadel Study Group. Gliadel wafer in initial surgery for malignant glioma: long-term follow-up of a multicenter controlled trial. Acta Neurochir. 148:269–275. 2006. View Article : Google Scholar : PubMed/NCBI
3	Sanai N, Polley MY, McDermott MW, Parsa AT and Berger MS: An extent of resection threshold for newly diagnosed glioblastomas. J Neurosurg. 115:3–8. 2011. View Article : Google Scholar : PubMed/NCBI
4	Lacroix M, Abi-Said D, Fourney DR, et al: A multivariate analysis of 416 patients with glioblastoma multiforme: prognosis, extent of resection, and survival. J Neurosurg. 95:190–198. 2001. View Article : Google Scholar
5	Fischer F, Dickson EF, Kennedy JC and Pottier RH: An affordable, portable fluorescence imaging device for skin lesion detection using a dual wavelength approach for image contrast enhancement and aminolaevulinic acid-induced protoporphyrin IX. Part II In vivo testing. Lasers Med Sci. 16:207–212. 2001. View Article : Google Scholar
6	Yamamoto J, Yamamoto S, Hirano T, et al: Monitoring of singlet oxygen is useful for predicting the photodynamic effects in the treatment for experimental glioma. Clin Cancer Res. 12:7132–7139. 2006. View Article : Google Scholar : PubMed/NCBI
7	Stummer W, Novotny A, Stepp H, Goetz C, Bise K and Reulen HJ: Fluorescence-guided resection of glioblastoma multiforme by using 5-aminolevulinic acid-induced porphyrins: a prospective study in 52 consecutive patients. J Neurosurg. 93:1003–1013. 2000. View Article : Google Scholar : PubMed/NCBI
8	Tamura Y, Kuroiwa T, Kajimoto Y, Miki Y, Miyatake S and Tsuji M: Endoscopic identification and biopsy sampling of an intraventricular malignant glioma using a 5-aminolevulinic acid-induced protoporphyrin IX fluorescence imaging system. Technical note. J Neurosurg. 106:507–510. 2007. View Article : Google Scholar : PubMed/NCBI
9	Della Puppa A, De Pellegrin S, d’Avella E, et al: 5-aminolevulinic acid (5-ALA) fluorescence guided surgery of high-grade gliomas in eloquent areas assisted by functional mapping. Our experience and review of the literature. Acta Neurochir. 155:965–972. 2013. View Article : Google Scholar : PubMed/NCBI
10	Shapiro WR, Green SB, Burger PC, et al: Randomized trial of three chemotherapy regimens and two radiotherapy regimens and two radiotherapy regimens in postoperative treatment of malignant glioma. Brain Tumor Cooperative Group Trial 8001. J Neurosurg. 71:1–9. 1989. View Article : Google Scholar : PubMed/NCBI
11	Laperriere N, Zuraw L and Cairncross G; Cancer Care Ontario Practice Guidelines Initiative Neuro-Oncology Disease Site Group. Radiotherapy for newly diagnosed malignant glioma in adults: a systematic review. Radiother Oncol. 64:259–273. 2002. View Article : Google Scholar : PubMed/NCBI
12	Mimura S, Ito Y, Nagayo T, et al: Cooperative clinical trial of photodynamic therapy with photofrin II and excimer dye laser for early gastric cancer. Lasers Surg Med. 19:168–172. 1996. View Article : Google Scholar : PubMed/NCBI
13	Mlkvy P, Messmann H, Pauer M, et al: Distribution and photodynamic effects of meso-tetrahydroxyphenylchlorin (mTHPC) in the pancreas and adjacent tissues in the Syrian golden hamster. Br J Cancer. 73:1473–1479. 1996. View Article : Google Scholar : PubMed/NCBI
14	Yamamoto J, Hirano T, Li S, et al: Selective accumulation and strong photodynamic effects of a new photosensitizer, ATX-S10. Na (II), in experimental malignant glioma. Int J Oncol. 27:1207–1213. 2005.PubMed/NCBI
15	Schaffer M, Ertl-Wagner B, Schaffer PM, et al: Feasibility of photofrin II as a radiosensitizing agent in solid tumors - preliminary results. Onkologie. 29:514–519. 2006. View Article : Google Scholar : PubMed/NCBI
16	Luksiene Z, Juzenas P and Moan J: Radiosensitization of tumours by porphyrins. Cancer Lett. 235:40–47. 2006. View Article : Google Scholar
17	Berg K, Luksiene Z, Moan J and Ma L: Combined treatment of ionizing radiation and photosensitization by 5-aminolevulinic acid-induced protoporphyrin IX. Radiat Res. 142:340–346. 1995. View Article : Google Scholar : PubMed/NCBI
18	Yamamoto J, Ogura S, Tanaka T, et al: Radiosensitizing effect of 5-aminolevulinic acid-induced protoporphyrin IX in glioma cells in vitro. Oncol Rep. 27:1748–1752. 2012.PubMed/NCBI
19	Yamamoto J, Ogura S, Shimajiri S, et al: 5-Aminolevulinic acid-induced protoporphyrin IX with multi-dose ionizing irradiation enhances host antitumor response and strongly inhibits tumor growth in experimental glioma in vivo. Mol Med Rep. (In press).
20	Riley PA: Free radicals in biology: oxidative stress and the effects of ionizing radiation. Int J Radiat Biol. 65:27–33. 1994. View Article : Google Scholar : PubMed/NCBI
21	Hall EJ and Giaccia AJ: Physics and chemistry of radiation absorption. Radiobiology for the Radiologist. Hall EJ and Giaccia AJ: 6th edition. Lippincott Williams & Wilkins; Philadelphia, PA: pp. 5–15. 2006
22	Chen Q, Chai YC, Mazumder S, et al: The late increase in intracellular free radical oxygen species during apoptosis is associated with cytochrome c release, caspase activation, and mitochondrial dysfunction. Cell Death Differ. 10:323–334. 2003. View Article : Google Scholar : PubMed/NCBI
23	Ogura A, Oowada S, Kon Y, et al: Redox regulation in radiation-induced cytochrome c release from mitochondria of human lung carcinoma A549 cells. Cancer Lett. 277:64–71. 2009. View Article : Google Scholar : PubMed/NCBI
24	Saenko Y, Cieslar-Pobuda A, Skonieczna M and Rzeszowska-Wolny J: Changes of reactive oxygen and nitrogen species and mitochondrial functioning in human K562 and HL60 cells exposed to ionizing radiation. Radiat Res. 180:360–366. 2013. View Article : Google Scholar : PubMed/NCBI
25	Yamamori T, Yasui H, Yamazumi M, et al: Ionizing radiation induces mitochondrial reactive oxygen species production accompanied by upregulation of mitochondrial electron transport chain function and mitochondrial content under control of the cell cycle checkpoint. Free Radic Biol Med. 53:260–270. 2012. View Article : Google Scholar : PubMed/NCBI
26	Ishizuka M, Abe F, Sano Y, et al: Novel development of 5-aminolevurinic acid (ALA) in cancer diagnoses and therapy. Int Immunopharmacol. 11:358–365. 2011. View Article : Google Scholar
27	Ogura S, Maruyama K, Hagiya Y, et al: The effect of 5-aminolevulinic acid on cytochrome c oxidase activity in mouse liver. BMC Res Notes. 4:662011. View Article : Google Scholar : PubMed/NCBI
28	Sugiyama Y, Hagiya Y, Nakajima M, Ishizuka M, Tanaka T and Ogura S: The heme precursor 5-aminolevulinic acid disrupts the Warburg effect in tumor cells and induces caspase-dependent apoptosis. Oncol Rep. 31:1282–1286. 2014.
29	Hosokawa Y, Sakakura Y, Tanaka L, Okumura K, Yajima T and Kaneko M: Radiation-induced apoptosis is independent of caspase-8 but dependent on cytochrome c and the caspase-9 cascade in human leukemia HL60 cells. J Radiat Res. 46:293–303. 2005. View Article : Google Scholar : PubMed/NCBI
30	Kowaltowski AJ, de Souza-Pinto NC, Castilho RF and Vercesi AE: Mitochondria and reactive oxygen species. Free Radic Biol Med. 47:333–343. 2009. View Article : Google Scholar : PubMed/NCBI
31	Le Caër S: Water radiolysis: influence of oxide surfaces on H₂ production under ionizing radiation. Water. 3:235–253. 2011. View Article : Google Scholar
32	Wallace SS: Enzymatic processing of radiation-induced free radical damage in DNA. Radiat Res. 150(Suppl 5): S60–S79. 1998. View Article : Google Scholar : PubMed/NCBI
33	Ward JF: DNA damage produced by ionizing radiation in mammalian cells: identities, mechanisms of formation, and reparability. Prog Nucleic Acid Res Mol Biol. 35:95–125. 1988. View Article : Google Scholar : PubMed/NCBI
34	Finkel T: Signal transduction by reactive oxygen species. J Cell Biol. 194:7–15. 2011. View Article : Google Scholar : PubMed/NCBI
35	Matoba S, Kang JG, Patino WD, et al: p53 regulates mitochondrial respiration. Science. 312:1650–1653. 2006. View Article : Google Scholar : PubMed/NCBI
36	Herrmann PC, Gillespie JW, Charboneau L, et al: Mitochondrial proteome: altered cytochrome c oxidase subunit levels in prostate cancer. Proteomics. 3:1801–1810. 2003. View Article : Google Scholar : PubMed/NCBI
37	Zhou S, Kachhap S and Singh KK: Mitochondrial impairment in p53-deficient human cancer cells. Mutagenesis. 18:287–292. 2003. View Article : Google Scholar : PubMed/NCBI
38	Warburg O: On respiratory impairment in cancer cells. Science. 124:269–270. 1956.PubMed/NCBI