OBJECTIVE: Photodynamic therapy has become an important method in clinical tumor treatment. This study aimed to investigate the effects of hematoporphyrin on multiple myeloma (MM) and its potential applications. METHODS: The MM cell line RPMI 8226 was treated with hematoporphyrin derivative (HPD), and CCK-8 assay was used to determine cell viability, apoptosis was detected by flow cytometry, intracellular reactive oxygen species (ROS) levels were measured using a detection kit combined with flow cytometry, and Western blot assay was used to detect apoptosis-related proteins and key signaling pathway protein levels. RESULTS: The optimal incubation time for the maximum absorption of HPD in RPMI 8226 cells was 4 hours. HPD significantly inhibited the proliferation of RPMI 8226 cells in a dose- and illumination time-dependent manner ( r =0.981; r =0.961). Additionally, HPD induced apoptosis in RPMI 8226 cells, but had no significant inhibitory effect on peripheral blood mononuclear cells derived from healthy individuals. HPD combined with illumination treatment significantly increased the intracellular ROS level, upregulated the expression of apoptosis-related proteins such as cleaved PARP, cleaved caspase-3 and Bax, and down-regulated the expression of proteins that maintain cell survival, such as NF-κB and Akt. CONCLUSION The HPD can inhibit the proliferation and induce apoptosis of multiple myeloma cells.
Humans ; Multiple Myeloma/pathology* ; Hematoporphyrins/pharmacology* ; Apoptosis/drug effects* ; Cell Line, Tumor ; Reactive Oxygen Species/metabolism* ; Cell Proliferation/drug effects* ; Photochemotherapy ; Cell Survival/drug effects* ; Signal Transduction

Hematoporphyrin monomethyl ether (HMME) combined with He-Ne laser irradiation is a novel and promising photodynamic therapy (PDT)-induced apoptosis that can be applied in vitro on canine breast cancer cells. However, the exact pathway responsible for HMME-PDT in canine breast cancer cells remains unknown. CHMm cells morphology and apoptosis were analyzed using optical microscope, terminal deoxynucleotidyl transferase dUTP nick end labeling fluorescein staining and DNA ladder assays. Apoptotic pathway was further confirmed by Real-time-polymerase chain reaction and Western blotting assays. Our results showed that HMME-PDT induced significant changes in cell morphology, such as formation of cytoplasmic vacuoles and the gradual rounding of cells coupled with decreased size and detachment. DNA fragmentation and cell death was shown to occur in a time-dependent manner. Furthermore, HMME-PDT increased the activities of caspase-9 and caspase-3, and released cytochrome c from mitochondria into the cytoplasm. HMME-PDT also significantly increased both mRNA and protein levels of Bax and decreased P53 gene expression in a time-dependent manner, while the mRNA and protein expression of Bcl-2 were repressed. These alterations suggest that HMME-PDT induced CHMm cell apoptosis via the mitochondrial apoptosis pathway and had anti-canine breast cancer effects in vitro.
Apoptosis* ; Blotting, Western ; Breast Neoplasms* ; Breast* ; Caspase 3 ; Caspase 9 ; Cell Death ; Cytochromes c ; Cytoplasm ; DNA ; DNA Fragmentation ; DNA Nucleotidylexotransferase ; Ether* ; Fluorescein ; Genes, p53 ; Hematoporphyrins* ; In Vitro Techniques ; Mitochondria ; Photochemotherapy ; RNA, Messenger ; Vacuoles

This investigation was made with regard to the influences of ultrasound combined with hematoporphyrin on the activities of antioxidative enzyme in ascites hepatoma 22 (H-22) tumor cells, and to a better understanding of the potential biological mechanism of sonodynamic therapy which involved the damage to cells. Combined with 100 microg/ml hematoporphyrin, high intensity focused ultrasound sonication at a frequency of 1.43 MHz and an intensity level of 2.0 W/cm2 was delivered to H-22 tumor cells for 1 min. The viability of cells was evaluated by typan-blue blue exclusion test. The intracellular reactive oxygen species (ROS) levels were determined by 2',7'-dichlorofluorescein diacetata (DCFH-DA). Enzymatic chemical methods were used to measure the activities of key antioxidative enzymes. The results indicated that the cell damage rate of ultrasound combined with hematoporphyrin was significantly higher than that of the treatment with ultrasound alone, and hematoporphyrin alone had no killing effect on H-22 cells. The level of ROS in cell suspension was significantly increased, and the key antioxidative enzyme activities were obviously decreased after treatment with the combined use of ultrasound and hematoporphyrin. We speculated that the decreased activities of key antioxidative enzymes in cells might be involved in mediating the killing effect on H22 cells in sonodynamic therapy.
Animals ; Female ; Glutathione Peroxidase ; metabolism ; Hematoporphyrins ; administration & dosage ; radiation effects ; Liver Neoplasms, Experimental ; enzymology ; therapy ; Mice ; Mice, Inbred ICR ; Photochemotherapy ; methods ; Photosensitizing Agents ; administration & dosage ; radiation effects ; Superoxide Dismutase ; metabolism ; Ultrasonics

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.
Biopsy ; Disulfiram ; Endosonography ; Esophageal Neoplasms* ; Follow-Up Studies ; Gastroscopy ; Hematoporphyrin Derivative ; Hematoporphyrins ; Humans ; Karnofsky Performance Status ; Lymph Nodes ; Male ; Neoplasm Metastasis ; Photochemotherapy* ; Quality of Life ; Recurrence

OBJECTIVETo investigate the killing effect of photodynamic therapy (PDT) mediated by hematoporphyrin derivative (HpD) on human colon carcinoma LoVo and CoLo205 cells in vitro.

METHODSLoVo and CoLo205 cells cultured in vitro were incubated in the presence of 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, and 4.0 microg/ml HpD for 4 h and exposed to different light doses delivered using a semiconductor laser at 630 nm with the energy density of 2, 5, 10, and 20 J/cm(2). After further culture for 24 h, the survival rate of LoVo and CoLo205 cells were analyzed by MTT assay, and the cellular fluorescence intensities of HpD were measured with a luminescence spectrometer.

RESULTSHpD-PDT resulted in effective cell killing to a comparable magnitude in LoVo and CoLo205 cells cultured in vitro (P>0.05). The killing effects were positively correlated with the concentration of HpD and the dosage of laser irradiation. Exposure to 20 J/cm(2) resulted in an IC(50) of LoVo and CoLo205 cells of 0.4 and 0.6 microg/ml respectively, which were not significantly different (P>0.05). The cellular HpD fluorescence intensities were also similar between the two cells.

CONCLUSIONHpD-PDT may effectively kill LoVo and CoLo205 cells cultured in vitro.

Animals ; Cell Line, Tumor ; Cell Survival ; drug effects ; radiation effects ; Colonic Neoplasms ; drug therapy ; pathology ; Dose-Response Relationship, Radiation ; Hematoporphyrins ; chemistry ; pharmacology ; Humans ; Lasers ; Photochemotherapy ; methods ; Photosensitizing Agents ; chemistry ; pharmacology ; Spectrometry, Fluorescence

OBJECTIVETo investigate the effect of HMME-PDT (Hematoporphyrin Monomethyl Ether-Photodynamic therapy) on Hyperplastic scar in the rabbit ear.

METHODSThe acute model of dermal Hyperplastic scar in the rabbit ear was established. 24 scars were randomly divided into 2 groups: the experimental group (n = 12) received HMME-PDT treatment, and the controlled group (n = 12) received no special treatment. Specimens were harvested from scars on postoperative 28 day. Scar hyper plasty and collagen fibers were observed by haematoxylin-eosin staining and Van-Gieson staining respectively. The microvessel density was calculated under microscope.

RESULTSCompared with the controlled group, HMME-PDT treatment in the experimental group reduced scar formation, decreased the microvessel density and prevented excess collagen deposition at the wound site.

CONCLUSIONSHMME-PDT may play a role in inhibiting hyperplastic scar in rabbit ear.

Animals ; Cicatrix, Hypertrophic ; pathology ; therapy ; Ear ; pathology ; Female ; Hematoporphyrins ; pharmacology ; Male ; Photochemotherapy ; Rabbits

OBJECTIVETo prepare photoimmunoconjugate of hematoporphyrin (HP) and herceptin, and study its killing and apoptosis-inducing effect on tumor cells BT-474.

METHODSHP-herceptin photoimmunoconjugate was synthesized with EDCI as the condensator. After exposure of the cells to 630 nm laser, the killing effect of the conjugate and cell apoptosis were evaluated by MTT assay and flow cytometry.

RESULTSCompared with free HP at equivalent dose, the immune reactivity, killing effect and the apoptosis-inducing effect of HP-herceptin immunoconjugate on BT-474 cells was enhanced (P<0.05).

CONCLUSIONThe killing effect of HP-herceptin immunoconjugate is stronger than free HP on BT-474 cells.

Antibodies, Monoclonal ; chemistry ; pharmacology ; Antibodies, Monoclonal, Humanized ; Antineoplastic Agents ; chemistry ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Survival ; drug effects ; Drug Compounding ; methods ; Flow Cytometry ; Hematoporphyrin Photoradiation ; methods ; Hematoporphyrins ; chemistry ; pharmacology ; Humans ; Immunoconjugates ; chemistry ; pharmacology ; Immunotherapy ; methods ; Photosensitizing Agents ; chemistry ; pharmacology ; Trastuzumab

PURPOSE: In order to elucidate the antitumor effect of photodynamic therapy (PDT), using a derivative of the photosensitizing agent hematoporphyrin (Photogem) and a diode laser, the cell death of uterine cancer cell lines (CaSki, HT3, HeLa, and SKOV-3), and mice transplanted with TC-1 lung cancer cells, were evaluated. MATERIALS AND METHODS: The morphological changes, MTT assay, flow cytometry, cytotoxicity and tumor growth inhibition study were evaluated at various time intervals after the PDT. RESULTS: The results showed that the survival rates of each cell line decreased with time and dose response after performing the PDT. Also, the PDT-induced damage of cancer cells was almost entirely confined to necrosis of the tumor cells in the early time courses. The irradiation of CaSki cells in the presence of Photogem induced plasma membrane disruption and cell shrinkage, indicating the plasma membrane as the main target for Photogem. In the in vivo experiment, significantly longer survival and a significantly smaller tumor size were seen over the time courses of the Photogem with irradiation compared to the untreated control groups; resorption of the tumor was also observed after the PDT treatment. CONCLUSION: Collectively, our results indicated that Photogem possesses anti-tumor effects, and necrosis-like death, with plasma membrane damage, was postulated to be the principal mechanism of the antitumor effect of the PDT using Photogem.
Animals ; Cell Death ; Cell Line* ; Cell Membrane ; Flow Cytometry ; Hematoporphyrins ; Lasers, Semiconductor ; Lung Neoplasms ; Mice ; Necrosis* ; Photochemotherapy ; Survival Rate ; Uterine Cervical Neoplasms* ; Uterine Neoplasms

Animal ; Antineoplastic Agents, Phytogenic/isolation & purification/*pharmacology ; Chlorophyll/*analogs & derivatives/isolation & purification/pharmacology/radiation effects ; Comparative Study ; Drug Screening Assays, Antitumor ; Feces/*chemistry ; Hematoporphyrin Derivative ; Hematoporphyrins/pharmacology ; Human ; Leukocytes, Mononuclear/drug effects ; Mice ; Oxygen/metabolism ; Photochemistry ; Photochemotherapy ; Radiation-Sensitizing Agents/isolation & purification/*pharmacology ; Silkworms/*metabolism ; Singlet Oxygen ; Spectrophotometry ; Structure-Activity Relationship ; Support, Non-U.S. Gov't ; Tumor Cells, Cultured/*drug effects/radiation effects

The mutagenic effect of HpD on cell SCE and the reactions of cell SCE to different sources of light combined with HpD were studied using V79 cells. There were 6 doses of HpD: 1 microgram/ml, 3 micrograms/ml, 5 micrograms/ml, 10 micrograms/ml, 50 micrograms/ml and 100 micrograms/ml. The dose of 5 micrograms/ml is equal to the maximum dose of HpD used in the clinic (HpD per milliliter of patient's blood). Our experiments demonstrated that when the cells were cultured in the dark and HpD was added to the medium no more than 5 micrograms/ml, the SCE frequencies were not increased. The cells were irradiated with different sources of light without HpD, both the fluorescence and ultraviolet light could promote SCE but the light of daylight lamp and red light did not increase it. But when HpD was added into culture medium at the dose of less than 5 micrograms/ml, every light could increase the cell SCE intensively except the daylight lamp light. The red light was more notable than the others by relation analysis.
Cells, Cultured ; Fluorescence ; Hematoporphyrin Photoradiation ; Hematoporphyrins ; pharmacology ; Humans ; Light ; Photochemotherapy ; Sister Chromatid Exchange ; drug effects ; Ultraviolet Rays