Identification of the targets of hematoporphyrin derivative in lung adenocarcinoma using integrated network analysis

BACKGROUND: Hematoporphyrin derivative (HPD) has a sensibilization effect in lung adenocarcinoma. This study was conducted to identify the target genes of HPD in lung adenocarcinoma. METHODS: RNA sequencing was performed using the lung adenocarcinoma cell line A549 after no treatment or treatment with X-ray or X-ray + HPD. The differentially expressed genes (DEGs) were screened using Mfuzz package by noise-robust soft clustering analysis. Enrichment analysis was carried out using "BioCloud" online tool. Protein-protein interaction (PPI) network and module analyses were performed using Cytoscape software. Using WebGestalt tool and integrated transcription factor platform (ITFP), microRNA target and transcription factor (TF) target pairs were separately predicted. An integrated regulatory network was visualized with Cytoscape software. RESULTS: A total of 815 DEGs in the gene set G1 (continuously dysregulated genes along with changes in processing conditions [untreated-treated with X-ray-X-ray + treated with HPD]) and 464 DEGs in the gene set G2 (significantly dysregulated between X-ray + HPD-treated group and untreated/X-ray-treated group) were screened. The significant module identified from the PPI network for gene set G1 showed that ribosomal protein L3 (RPL3) gene could interact with heat shock protein 90 kDa alpha, class A member 1 (HSP90AA1). TFs AAA domain containing 2 (ATAD2) and protein inhibitor of activated STAT 1 (PIAS1) were separately predicted for the genes in gene set G1 and G2, respectively. In the integrated network for gene set G2, ubiquitin-specific peptidase 25 (USP25) was targeted by miR-200b, miR-200c, and miR-429. CONCLUSION: RPL3, HSP90AA1, ATAD2, and PIAS1 as well as USP25, which is targeted by miR-200b, miR-200c, and miR-429, may be the potential targets of HPD in lung adenocarcinoma.

Research progress of the anti-tumor effect of sonodynamic and photodynamic therapy / 生物医学工程学杂志

Cancer, as a serious threat to human health, is one of the major killers. The treatment of cancer has attracted more and more attention. Currently, the means of treating cancer is also increasing, but there is no emergence of a fully satisfactory treatment. A combination of sonodynamic therapy (SDT) and photodynamic therapy (PDT), named sono-photodynamic therapy (S-PDT), is a new composite cancer therapy. Because the therapy can significantly improve the tumor curing effect, it has good application prospects in cancer prevention and treatment. The present article reviewed the progress of the anti-tumor mechanisms and influencing factors of S-PDT.

Avaliação do efeito citotóxico da terapia fotodinâmica associada ao LED e ao Photogem® sobre a mucosa bucal de rato / Toxicity of the photodynamics therapy with LED associated to Photogem®: an in vivo study

A utilização da PDT para tratamento de diferentes tipos de infecções, tal como a candidose bucal, tem sido estudada. Entretanto, poucos são os dados científicos que relatam os possíveis efeitos tóxicos dessa terapia. Dessa forma, o objetivo deste estudo foi avaliar os efeitos da irradiação na mucosa bucal de ratos com LED azul (de 460 nm e potência de 200 mW/cm2) em presença do fotossensibilizador (FS) Photogem®, em duas diferentes concentrações (500 mg/L e 1000 mg/L). Para isso, foram utilizados 101 ratos (Rattus Norvegicus Albinus Holtzman) distribuídos em 6 grupos, de acordo com os seguintes tratamentos: Grupo 1 ­ controle; Grupo 2 ­ aplicação do FS (500 mg/L); Grupo 3 ­ aplicação do FS (500 mg/L) e irradiação com LED; Grupo 4 - aplicação do FS (1000 mg/L); Grupo 5 ­ aplicação do FS (1000 mg/L) e irradiação com LED; e Grupo 6 ­ irradiação com LED. O FS foi aplicado por 30 minutos (tempo de pré-incubação) e o tempo de irradiação da mucosa foi de 20 minutos (dose de 144 J/cm2 ). Decorridos os 4 períodos de avaliação propostos (0 dia, 1dia, 3 dias e 7 dias), os animais tiveram a mucosa palatina fotografada para análise macroscópica, sendo então imediatamente sacrificados para remoção cirúrgica do palato e posterior análise em microscopia de luz e de fluorescência. Um mapeamento térmico foi realizado a fim de avaliar a variação de temperatura ocorrida no tecido durante a irradiação com LED. Macroscopicamente, em todos os grupos experimentais e para todos os períodos de avaliação propostos na presente pesquisa, observou-se que a mucosa apresentava-se intacta, com aspecto de normalidade semelhante ao do Grupo 1 (controle). Microscopicamente, alterações teciduais, caracterizadas especialmente por discreta inflamação, puderam ser observadas na mucosa palatina de apenas 4 de um total de 80 animais submetidos a PDT. A penetração do fotossensibilizador na mucosa tratada pôde ser observada por meio da emissão de fluorescência do Photogem® , tendo este FS se mantido presente apenas no tecido epitelial. O mapeamento térmico revelou que a temperatura aumentou de 35ºC para 41ºC durante 20 minutos de irradiação. Dentro das condições experimentais avaliadas, foi possível concluir que a PDT, utilizando Photogem® nas concentrações de 500 mg/L e 1000 mg/L associado ou não à irradiação com LED (dose de 144 J/cm2), não foi tóxica para a mucosa palatina de ratos

The use of PDT has been investigated for the treatment of different types of infection, like oral candidosis. There are, however, few research-based data that report the possible toxic effects of this therapy. Therefore, this study evaluated the effects of irradiating the palatal mucosa of rats with blue LED (460 nm; 200 mW/cm²) in the presence of the photosensitizer Photogem® at two concentrations (500 and 1000 mg/L). Then, 101 rats (Rattus norvegicus albinus Holtzman) were randomly distributed in six groups, according to the treatment performed on the palatal mucosa: Group 1: control; Group 2: Photogem® (500 mg/L); Group 3: Photogem® (500 mg/L) + blue LED; Group 4 - Photogem® (1000 mg/L); Group 5: (1000 mg/L) + blue LED; and Group 6: blue LED. The exposure times to the photosensitizing agent and to the light source were 30 min (pre-incubation time) and 20 min (144 J/cm2 energy density), respectively. At 0, 1, 3 and 7 days posttreatment, the animals had their palatal mucosa photographed for macroscopic analysis and were immediately sacrificed. The palate was removed for further analysis by light and fluorescence microscopy. Thermal mapping was made to evaluate the temperature change occurred in the tissue during LED irradiation. In all experimental groups and periods, the macroscopic analysis revealed intact mucosa with normal aspect similar to that of Group 1 (control). Tissue alterations, characterized primarily by a mild inflammation, were observed microscopically on the mucosa of only 4 out of 80 animals subjected to PDT. Photosensitizer penetration into the treated mucosa was identified by the fluorescence emitted by Photogem® and was limited to the epithelial layer. The thermal mapping revealed a temperature increase from 35 to 41ºC during the 20-min irradiation. In conclusion, under the tested conditions, PDT using Photogem® at 500 and 1000 mg/L concentrations associated or not to LED irradiation (144 J/cm2) was not toxic to the rat palatal mucosa

Efeito citotóxico da terapia fotodinâmica associando Photogem® e LED azul e vermelho em cultura de células normais / Cytotoxic effect of the photodynamic therapy associating Photogem® and blue or red LED on normal cell culture

Para considerar a Terapia Fotodinâmica (PDT) como tratamento clínico da estomatite protética, é necessário conhecer tanto o potencial antifúngico, como efeito citotóxico desta terapia sobre células normais do indivíduo. Assim, o objetivo desse estudo in vitro foi avaliar a citotoxicidade da PDT antifúngica com o fotossensibilizador Photogem® associado ao LED azul e ao LED vermelho em cultura de fibroblastos L929 e células odontoblastóides MDPC-23. As células foram cultivadas (30.000 células/cm2) em placas de 24 compartimentos por 48 horas e ambos os tipos celulares foram incubados com Photogem® (0, 10, 25, 50, 100 ou 150 mg/L) e irradiados ou não pelo LED azul (460 ± 3 nm; 25,5 ou 37,5 J/cm2; 22 mW/cm2) ou LED vermelho (630 ± 3 nm; 70 ou 100 J/cm2; 25 mW/cm2) . O metabolismo celular foi determinado 0, 12 e 24 horas após a PDT utilizando o teste do metiltetrazolium (MTT), e a morfologia celular avaliada pela microscopia eletrônica de varredura (MEV). A técnica de citometria de fluxo foi utilizada para avaliar o tipo de morte celular (necrose ou apoptose) assim como estimar os níveis intracelulares das espécies reativas de oxigênio (EROs). Observou-se redução do metabolismo celular estatisticamente significante para todas as concentrações do Photogem® quando irradiadas em qualquer dose de luz, sendo essa redução de 90 a 97% tanto para as células L929 quanto MDPC-23 (ANOVA and Dunnet's post hoc tests; p<0.05). Essa redução da atividade mitocondrial não foi dependente da concentração do fotossensibilizador e nem da dose de luz empregada. Também, foi demonstrado que a atividade mitocondrial das células submetidas a PDT não foi recuperada após 12 ou 24 horas, caracterizando um dano irreversível. A presença do Photogem® e da luz isoladamente não alterou estatisticamente a atividade mitocondrial de ambas as linhagens. As células submetidas à PDT tiveram sua morfologia alterada, não sendo possível observação dos limites celulares. Em ambas as linhagens, foi observado predomínio de morte celular por necrose quando em contato com o fotossensibilizador. A presença do Photogem® e a exposição à luz aumentaram os níveis de EROs intracelulares de uma forma dosedependente para L929 e MDPC-23. A associação do Photogem® com LED azul ou vermelho causou intensos efeitos tóxicos sobre cultura de células normais, caracterizados pela redução da atividade mitocondrial, alterações morfológicas e indução de morte celular por necrose

In order to consider the photodynamic therapy (PDT) as a clinical treatment for candidosis, it is necessary to know its antifungal potential and its cytotoxicity effect on normal cells. Therefore, the purpose of this in vitro study was to evaluate the cytotoxicity of PDT with Photogem® associated to blue and red LED on L929 and MDPC-23 cell cultures. The cells (30.000 cells/cm2) were seeded in 24-well plates for 48 hours, incubated with Photogem® (10, 25, 50, 100 or 150 mg/L) and were irradiated or not with a blue LED source (460 ± 3 nm; 25.5 or 37.5 J/cm2; 22 mW/cm2) or with a red LED source (630 ± 3 nm; 75 or 100 J/cm2; 22 mW/cm2). Cell metabolism was evaluated by the MTT assay (ANOVA and Dunnet's post hoc tests; p<0.05) and cell morphology was examined by scanning electron microscopy. Flow cytometry was employed to analyze the type of PDT-induced cell death (necrosis or apoptosis) as well as to estimate intracellular production of reactive oxygen species (ROS). There was a statistically significant decrease of mitochondrial activity for all Photogem® concentrations associated to blue or red LED regardless irradiation time; this reduction ranged from 90 to 97% for both cell lines. This reduction, however, was not dependent on the photosensitizer concentration. It was also demonstrated that the mitochondrial activity of the cells submitted to PDT was not recovered after 12 or 24 hours, characterizing irreversible cell damage. PDT-treated cells presented an altered morphology with ill-defined limits. In both cell lines, there was a predominance of necrotic cell death when in contact with the photosensitizer. The presence of Photogem® and exposure to blue and red LED increased the intracellular levels of ROS in both L929 and MDPC23 cells in a dose-dependent manner. The association of Photogem® and blue LED caused severe toxic effects on normal cell culture, characterized by the reduction of the mitochondrial activity, morphological alterations and induction of necrotic cell death

Mechanism of photodynamic therapy against human esophageal carcinoma xenografts in nude mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the mechanism of photodynamic therapy (PDT) in nude mice bearing human esophageal cancer cell line Eca-109 xenografts.</p><p><b>METHODS</b>A nude mouse model bearing human esophageal carcinoma was established by subcutaneous transplantation of Eca-109 cells. The mice were then randomized into 4 groups, namely hematoporphyrin derivative (HpD)-PDT group (given HpD and laser irradiation), exclusive laser irradiation group, exclusive HpD group and blank control group. In HpD-PDT group, the mice were exposed to irradiation at the light energy density of 120 Jsol;cm(2) delivered via a DIOMED 630 PDT system 24 h after intraperitoneal HpD injection, and the mice in exclusive laser irradiation group received only laser irradiation. Three days later, all the nude mice were sacrificed for determination of malondialdehyde (MDA) production, immunohistochemistry for caspase-3 protein and HE staining of the tumor tissue.</p><p><b>RESULTS</b>The MDA level was significantly higher in HpD-PDT group than in the other 3 groups (P<0.01), and comparable between the latter 3 groups. Expression of caspase-3 protein was similar between HpD-PDT group and the blank control group (P>0.05). Under light microscope, HE staining visualized massive tissue necrosis in HpD-PDT group with homogeneous red staining.</p><p><b>CONCLUSION</b>In human esophageal carcinoma xenografts in nude mice, HpD-PDT generates singlet oxygen to result in direct tumor cell damage and cause MDA production. Caspase-3 may not be activated in the apoptotic pathway, suggesting that this pathway may not be caspase-3-dependent.</p>

Main factors affecting the effect of photodynamic therapy against human esophageal cancer cells in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the tumor cell-killing effect of photodynamic therapy against human esophageal cancer cells in vitro and identify the main factors affecting the effect.</p><p><b>METHODS</b>Human esophageal cancer Eca-109 cells were incubated for 24 h in vitro with hematoporphyrin derivative (HpD) and Photofrin at different concentrations prior to exposure to a light energy density of 15 J/cm(2) delivered from a DIOMED 630 PDT system. The cell killing effect was also evaluated for different HpD concentrations combined with 3 light energy densities (10, 30, and 50 J/cm(2)), respectively. The cell survival rate was measured using MTT assay, and fluorescence spectrometry was used to detect the intracellular photosensitizer fluorescence of the tumor cells after incubation with HpD for 4 h.</p><p><b>RESULTS</b>The cell survival rate after incubation with the two photosensitizers at different concentrations were significantly different, and under the 3 different light energy densities, incubation of the cells with different HpD concentrations also resulted in significantly different cell survival rates (P<0.05). At the 4 low photosensitizer concentrations and with different light energy densities, the cell survival rates were similar (P>0.05), but the 4 higher photosensitizer concentrations resulted in significant difference in the cells survival (P<0.05). Correlation analysis showed that the intracellular photosensitizer concentration was positively correlated to the photosensitizer concentrations in cell incubation (r=0.997).</p><p><b>CONCLUSION</b>When the light source remains constant, the light energy density, the kinds of photosensitizers and their concentrations are the main factors affecting the Eca-109 cell-killing effect of PDT.</p>

Outcome of Photodynamic Therapy for Early Esophageal Cancer

BACKGROUND/AIMS: Endoscopic treatment as an alternative to surgery has become increasingly popular for improving the quality of life. Although photodynamic therapy (PDT) has been used for the endoscopic treatment of digestive cancer, its curative efficacy remains unclear. We evaluated the curative efficacy of PDT in superficial esophageal cancer in inoperable patients. METHODS: Ten male patients with histologically proven early esophageal cancer (surgery was contraindicated for age > 80 years, surgery was contraindicated, Karnofsky performance status of at least 30%, or refusal of surgery) were intravenously injected with a hematoporphyrin derivative (2 mg/kg), and PDT was performed 48 h later. The response to treatment was assessed by gastroscopy with biopsies. RESULTS: The mean follow-up period was 27.6 months (range, 9.6-58.7 months). Endoscopic ultrasonography revealed that all ten cases were at tumor stage T1. Complete remission (CR) to initial and subsequent PDT was observed in all patients. For the CR cases, the recurrence rate was 10% (1/10) and the time from initial PDT to recurrence was 9.6 months. CONCLUSIONS: For patients in whom surgery is risky or refused, PDT may represent an acceptable alternative treatment modality, especially for superficial esophageal cancer without lymph node metastasis. However, a study involving long-term follow-up in a large population is needed for confirmation.

Hematoporphyrin derivative-mediated photodynamic therapy for human nasopharyngeal carcinoma: a comparative study with CNE2 and C666-1 cell lines in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate biological effect of hematoporphyrin derivative (HpD) photodynamic therapy (PDT) on in vitro cultured nasopharyngeal carcinoma (NPC) cell lines CNE2 and C666-1.</p><p><b>METHODS</b>CNE2 and C666-1 cells cultured in vitro were incubated in a medium containing HpD at different concentrations (0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, and 4.0 microg/ml) for 4 h followed by exposure to different light doses (2, 5, 10, and 20 J/cm2) using a diode laser at 630 nm with power density of 20 mW/cm2. After 24 h of incubation with HpD-PDT, the survival rate of CNE2 and C666-1 cells were analyzed by MTT assay.</p><p><b>RESULTS</b>HpD-PDT produced effective killing of CNE2 and C666-1 cells cultured in vitro, and the killing effects were positively correlated with HpD concentration and the irradiation dose. Exposure of CNE2 and C666-1 cells to irradiation dose of 20 J/cm2 resulted in the IC50 of 0.7 and 1.2 microg/ml, respectively (P<0.01). With the same HpD concentration and irradiation dose, the survival rate of C666-1 cells, however, was significantly higher than that of CNE2 cells (P<0.05).</p><p><b>CONCLUSION</b>HpD-PDT may result in effective killing of CNE2 and C666-1 cells cultured in vitro, although C666-1 cells are less sensitive to HpD-PDT than CNE2 cells.</p>

Photodynamic Therapy for Superficial Gastrointestinal Tumors / 대한소화기내시경학회지

BACKGROUND/AIMS: Although photodynamic therapy (PDT) has been used for the endoscopic treatment of digestive cancer, its curative efficacy remains uncertain. This study evaluated the curative role of PDT in superficial gastrointestinal cancer. METHODS: Fifteen lesions in 14 patients with a histologically proven carcinoma (early esophageal cancer 6, early gastric cancer 8, ampulla of Vater cancer 1) were injected with an intravenous hematoporphyrin derivative (2 mg/kg), and PDT was performed 48 hours later. The response to treatment was assessed by gastroscopy with biopsies. RESULTS: The median follow-up time was 273 days (42~1,030 days). According to the TNM stage of endoscopic ultrasonography, there were 14 T1 cases and 1 T2 case. Complete remission was observed in 13 cases after the initial and consecutive PDT. There were 2 cases of failure. The recurrence rate was 15.4% (2/13), and the median time from the initial PDT to recurrence was 349 days. CONCLUSIONS: PDT using a hematoporphyrin derivative as a photosensitizer is a safe and efficient method for treating early cancer. However, a long-term follow up period using a large population sample will be needed for confirmation.

Photodynamic diagnosis and fluorescence-guided resection of malignant gliomas: a report of 15 cases / 中华外科杂志

<p><b>OBJECTIVE</b>To study the usefulness of the intraoperative photodynamic diagnosis (PDD) and fluorescence-guided resection of malignant gliomas.</p><p><b>METHODS</b>Fifteen consecutive patients with malignant gliomas received doses of hematoporphyrin derivative (HPD, 2 mg/kg body weight) 48 hours before induction of anesthesia. After the tumors recognized by bare eyes they were removed routinely. The fluorescence around 690 nm excited by laser beam (wavelength 632.5 nm) was detected by laser electronic spectrum analyzer and then fluorescing tissue was removed whenever it was considered safel. Tissue samples derived from the walls of tumor cavities after resection and PDD were sent for histological examination. Compared with the result of the histological examination, the sensitivity and specificity of PDD were calculated and recorded. Early postoperative MRI or CT were done to determine the extend of the resection of the tumors. Surgical mortality and morbidity were also recorded.</p><p><b>RESULTS</b>Intraoperatively, in all of 15 cases tumor areas with HPD fluorescence could be recognized by laser electronic spectrum analyzer. On the basis of 106 tissue samples derived from 15 tumors, a sensitivity of 90.6%, a specificity of 96.8% and an accuracy of 94.3% of PDD were achieved. In 2 cases the resection of residual tumor were performed after finding left tumors by PDD. Complete resection of contrast-enhancing tumor was accomplished in 9 patients (60%). Residual intraoperative tissue fluorescence left unresected for safety reasons predicted residual enhancement on MR images in 5 of the 6 remaining patients. No perioperative deaths and one case of morbidity were encountered.</p><p><b>CONCLUSIONS</b>Intraoperative photodynamic diagnosis following resection of malignant gliomas can detect residual tumor tissue with high accuracy. Photodynamic diagnosis and fluorescence-guided resection of malignant gliomas have a positive role in improving the radicality of malignant glioma resection.</p>

Experimental interstitial photodynamic therapy of the Morris 7777 hepatoma in the rat / 대한내과학회지

BACKGROUND: Photodynamic therapy (PDT) of hepatic tumors has been restricted due to the preferential retention of photosensitizers in normal liver tissue. Therefore, superficial tumor illumination causes subsequential liver necrosis. Moreover, the limited light penetration during superficial illumination makes it impossible to treat deep-seated or larger solid tumors. These limitations were overcome by interstitial therapy. Aim: The aim of study is to investigate the macroscopic and microscopic effect of a single session of interstitial photodynamic therapy on liver tumor. METHODS: The Morris 7777 hepatoma cells were injected subcutaneously into the flanks of Buffalo female rats. Tumor growth was monitored with measuring the tumor size. Animals were pretreated with hematoporphyrin derivative 48 hours prior to the intratumoral delivery of laser radiation, when the tumor had reached a volume greater than 1.0 cm3. One or two weeks after interstitial PDT, the extent of tumor necrosis was evaluated microscopically. RESULTS: Volume of lesions averaged 10.2 mm3. Histologic stains demonstrated microvascular thrombosis and coagulative necrosis within the lesions. There appeared to be 100% cellular destruction within the lesion by cytochemical staining. CONCLUSION: The results of this study show the ability of interstitial PDT to cause destruction of liver tumor.

Photodynamic therapy and its application in gynecologic oncology / 中国医学科学院学报

While photodynamic therapy is applied on neoplasm, photosensitisers tend to accumulate in neoplastic tissues. With appropriate wavelength light, it causes photochemical reaction and destructs neoplastic tissues. Its better selection for tumor tissue with effective photochemical reaction, and lower side effect make it widespread application in gynecologic oncology. At present, photodynamic therapy has been used in diagnosing and treating lower genital tract carcinoma in situ, and advanced malignant tumor such as vulval and ovarian carcinoma.

A Case of Bowen's Disease Partially Responded to Photodynamic Therapy

Photodynamic therapy(PDT) is a treatment modality by highly reactive oxygen intermediates generated through the interaction of light with a photosensiziter. It has been shown to be an effective treatment for various cutaneous and noncutaneous malignancies. It is efficient for the curative and palliative treatment of epithelial skin tumor in situ or early invasive lesions. In effect, it is a useful alternative treatment for the lesions located on anatomically difficult areas or the large-sized lesions. We treated a case of Bowen's disease arising on the plantar area and 3rd and 4th toewebs of left forefoot in a 61-year-old man with PDT using the hematoporphyrin derivative, porfirmer sodium(Photofrin, Russia) as a photosensitizer and gold vapor laser as a visible light source. The outcome showed partial clinical improvement after about 2 months' follow-up.

Effects of photodynamic treatment on biological antioxidant systems in rats.

Effects of photodynamic treatments on inherent antioxidant metabolites and cellular defence enzymes have been investigated in rats. Wistar rats were grouped into untreated controls, light controls, hematoporphyrin derivative (Hpd) (treated with 5 and 10 mg Hpd/kg body weight and kept in dark) and sets treated with both Hpd and red light (dose 172 and 344 j/m2 ). After 2, 24, 48 and 72 hr of Hpd injection the rats sacrificed, livers quickly excised to analyze Hpd uptake, activities of enzymes like catalase, GSH-Px and antioxidants like GSH, vitamin A, vitamin E and vitamin C. The results showed that the loss of Hpd from liver as a function of post- injection time was non- linear. An increased generation of lipid radicals was observed in the groups treated with 5 mg Hpd and higher dose of light and in groups treated with 10 mg Hpd at both the doses of light. Combination of light and Hpd reduced hepatic GSH content with a concomitant reduction in GSH-Px. At higher doses of Hpd and light, there was a significant reduction in hepatic vitamin A levels. Combination of Hpd and light in all doses reduced vitamin E content in liver. The decreased biological antioxidant contents and GSH-Px may be attributed to their utilization for the scavenging of free radicals generated by Hpd and light in tissues. However, no change in catalase activity and vitamin C content in liver was noted in experimental rats. The results suggest that exposure to higher doses of Hpd with light alters oxidant stress system and TBARS content in rat.

Clinical Experiences of Photodynamic Therapy in the Extramammary Paget's Disease / 대한피부과학회지

BACKGROUND: Photodynamic therapy(PDT) is a kind of non-invasive photochemotherapy, which is designed to kill the target cells through photodynamic reaction. It has been reported that PDT is very effective in superficial lesions, including extramammary Paget's disease. Yet, there has been no clinical trial for the treatment of extramammary Paget's disease using PDT in Korea. OBJECTIVE: The purpose of this study was to evaluate the effectiveness of PDT in treating extramammary Paget's disease METHOD: Seven patients with anogenital extramammary Paget's disease were treated repeatedly with local PDT using topical ALA 20% and intralesional injection of hematoporphyrin derivative (HpD), and systemic PDT using HpD. Fluences of 125-200J/cm2 and intensities of 50-100mW/cm2 were used. RESULT: The clinical outcomes were disappointing. All the patients experienced only partial responses despite repeated treatments. But side effects were minimal. CONCLUSION: PDT is not effective enough for the treatment of extramammary Paget's disease to be used as a primary therapy. It would be better to use as an adjuvant to other treatment methods. Photodynamic sensitization, also known as photodynamic diagnosis, might help us define a more reliable border of the extent involved and the surgical margin to be excised.

Clinical Experiences of Photodynamic Therapy in the Extramammary Paget's Disease / 대한피부과학회지

BACKGROUND: Photodynamic therapy(PDT) is a kind of non-invasive photochemotherapy, which is designed to kill the target cells through photodynamic reaction. It has been reported that PDT is very effective in superficial lesions, including extramammary Paget's disease. Yet, there has been no clinical trial for the treatment of extramammary Paget's disease using PDT in Korea. OBJECTIVE: The purpose of this study was to evaluate the effectiveness of PDT in treating extramammary Paget's disease METHOD: Seven patients with anogenital extramammary Paget's disease were treated repeatedly with local PDT using topical ALA 20% and intralesional injection of hematoporphyrin derivative (HpD), and systemic PDT using HpD. Fluences of 125-200J/cm2 and intensities of 50-100mW/cm2 were used. RESULT: The clinical outcomes were disappointing. All the patients experienced only partial responses despite repeated treatments. But side effects were minimal. CONCLUSION: PDT is not effective enough for the treatment of extramammary Paget's disease to be used as a primary therapy. It would be better to use as an adjuvant to other treatment methods. Photodynamic sensitization, also known as photodynamic diagnosis, might help us define a more reliable border of the extent involved and the surgical margin to be excised.

Cellular accumulation and biological activity of hematoporphyrin derivative(L) in comparison with photofrin II.

Hematoporphyrin derivative, a drug used in the photodynamic therapy of solid tumours was synthesized in the laboratory and was called Hpd(L). Physico-chemical and biological properties of this drug have been compared with Photofrin II, the commercially available drug. Both Hpd(L) and Photofrin II possess similar properties qualitatively. Quantitatively, Hpd(L) was half as active as Photofrin II in its efficacy in causing photodynamic cytotoxicity or in the optical densities at the absorption peaks. These differences could be due to the differences in the compositions. Hpd(L) is a non-purified complex mixture of a number of porphyrin derivatives whereas Photofrin II is a relatively purer compound consisting of di- and tri-hematoporphyrins linked through ether or ester bonds. In vitro cellular uptake and retention of these drugs has been found to be a passive process not involving energy expenditure. pH and temperature of the incubation media have been found to profoundly influence these processes, while a complex relation seems to exist between physiological state of a cell and accumulation of these photosensitizers.

Effect of hematoporphyrin derivative and light on sulfhydryl groups in brain tumour cells.

Effect of hematoporphyrin derivative (HpD) and light on sulfhydryl (SH) groups in brain tumor cells was studied. Sulfhydryl groups were measured by 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB) and a fluorescent probe 3,4-maleimidylphenyl-4-methyl-7-diethylaminocoumarin (CPM). Incubation of cells with HpD in dark resulted in the loss of DTNB as well as CPM reactive SH groups. After 2 hr of incubation DTNB reactive SH groups showed a negligible change while a continuous decrease was observed in CPM reactive SH groups. Cells treated with HpD showed a further degradation of SH groups upon light irradiation. A comparison of cytotoxicity and SH groups under identical conditions showed that blockage of SH groups by HpD binding is not leathal to the cells where as photoinduced cell death was observed on photodegradation of SH groups.

Structural alterations induced by photodynamic action of hematoporphyrin derivative (HpD) in plasma membrane of glioblastoma (U-87MG) cells: time dependent fluorescence spectroscopic study.

Photodynamic action of hematoporphyrin derivative (HpD) on the plasma membrane of human glioblastoma U-87MG cells was investigated using lipid and protein specific fluorescent probes trimethylammonium-1,6-diphenyl 1,3,5-hexatriene (TMA-DPH) and N-(1-pyrene)-maleimide (PM) respectively. Steady state anisotropy, decay time and time dependent anisotropy of these probes in U-87MG cells were measured. Light irradiation caused an increase in the steady state anisotropy of TMA-DPH in cells treated with HpD; however, no change in decay time was observed. Time dependent anisotropy measurements were performed and the data were analyzed using wobbling in cone model. A decrease in the rotational relaxation time (phi) as well as the cone angle (theta(c)) and an increase in the order parameter (S) of TMA-DPH were observed on photosensitization of cells. A decrease in the order parameter (S) of TMA-DPH were observed on photosensitization of cells. A decrease in the steady rate anisotropy and the rotational relaxation time (phi) of PM and enhancement in the lipid peroxidation were also observed. Our results show that the photodynamic action of HpD increases the order in the lipid bilayer and the mobility of the proteins in the plasma membrane of cells.