Cancer Res Treat.  2004 Dec;36(6):389-394.

Photodynamic Effects of Radachlorin(R) on Cervical Cancer Cells

• Bae SM
• Kim YW
• Lee JM
• Namkoong S
• Han SJ
• Kim JK
• Lee CH
• Chun HJ
• Jin HS
• Ahn WS

Abstract

PURPOSE
Photodynamic therapy (PDT) is a novel treatment modality, which produces local tissue necrosis with laser light following the prior administration of a photosensitizing agent. Radachlorin(R) has recently been shown to be a promising PDT sensitizer. In order to elucidate the antitumor effects of PDT using Radachlorin(R) on cervical cancer, growth inhibition studies on a HPV-associated tumor cell line, TC-1 cells in vitro and animals with an established TC-1 tumor in vivo were determined. MATERIALS AND METHODS: TC-1 tumor cells were exposed to various concentrations of Radachlorin(R) and PDT, with irradiation of 12.5 or 25 J/cm2 at an irradiance of 20 mW/cm2 using a Won-PDT D662 laser at 662 nm in vitro. C57BL/6 mice with TC-1 tumor were injected with Radachlorin(R) via different routes and treated with PDTin vivo. A growth suppression study was then used to evaluate the effects at various time points after PDT. RESULTS: The results showed that irradiation of TC-1 tumor cells in the presence of Radachlorin(R) induced significant cell growth inhibition. Animals with established TC-1 tumors exhibited significantly smaller tumor sizes over time when treated with Radachlorin(R) and irradiation. CONCLUSION: PDT after the application of Radachlorin(R) appears to be effective against TC-1 tumors both in vitro and in vivo.

Keyword

Radachlorin(R); Photodynamic therapy (PDT); Cervical cancer; TC-1 cell

MeSH Terms

Animals
Cell Line, Tumor
Mice
Necrosis
Photochemotherapy
Uterine Cervical Neoplasms*

Figure

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  Fig. 1 Structure of Radachlorin®'s major component - sodium chlorin e6 (A) and one of the minor components - sodium chlorin e6 (B).

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  Fig. 2 Development of Radachlorin®-mediated cellular uptake in TC-1 cells. Radachlorin® was incubated at concentrations of 0µg/ml (A), 2.5µg/ml (B), 5µg/ml (C), 10µg/ml (D), 20µg/ml (E) and 50µg/ml (F), for 24 h. (Magnification ×400 for all photographs).

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  Fig. 3 Intracellular localization of TC-1 cells after 12 h of exposure to 5µg/ml Radachlorin®, as measured by confocal microscopy (A-B) (Magnification×12,000). The fluorescence was emitted from well-defined spots in the cytoplasm, and diffused fluorescence seen in the entire cytoplasm.

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  Fig. 4 Cell growth-inhibitory effects of PDT on TC-1 cells in vitro. Cells (2×103 cells/well) were cultured overnight in 96-well plates, in triplicate, and incubated with Radachlorin® for 3 or 12 h, with irradiation of 12.5 or 25 J/cm2 at an irradiance of 20 mW/cm2 using a Won-PDT D662 laser at 662 nm. After PDT, the cells were cultured for a predetermined time, and the MTT assay performed. The conditions for the TC-1 cells were (A) Radachlorin® 3h incubation, and irradiation 25 J, 20 mW (B) 12 h, and 25 J, 20 mW (C) 3 h, and 12.5 J, 20 mW (D) 12 h, and 12.5 J, 20 mW; ●, Control; ○, 2.5µg/ml of Radachlorin®; ■, 5µg/ml of Radachlorin®; □, 10µg/ml of Radachlorin®; ▲, 20µg/ml of Radachlorin®.

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  Fig. 5 Antitumor effect of PDT using Radachlorin® in vivo. Tumor growth curves of mice treated using several protocols. (◆), control (untreated); (Δ), Radachlorin® only; (▲), irradiation only; (○), Radachlorin® 40 mg/kg b.w. i.v. injection and irradiation; (●), Radachlorin® 40 mg/kg b.w. i.p. injection and irradiation.

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