hypocrellin-b and Carcinoma

Hypocrellin B, a natural pigment from a traditional Chinese herb, has been attracting extensive attention. The present study aims to investigate whether hypocrellin B can enhance cell death induced by ultrasound sonification on nasopharyngeal carcinoma cells in vitro. The sonodynamic action of hypocrellin B was investigated on nasopharyngeal carcinoma cell line CNE2 cells as tumor model cells. In the experiments, the hypocrellin B concentration was kept constant at 2.5 microM and the cells were subject to ultrasound exposure for 15 s at an intensity of 0.65 W/cm(2). Cytotoxicity was investigated 24 h after ultrasound sonification. Apoptosis was evaluated using flow cytometry with annexin V-FITC and propidium iodine staining and nuclear staining with Hoechst 33258. Cell ultrastructure morphology was observed using transmission electron microscopy (TEM). No significant dark cytotoxicity of hypocrellin B in the CNE2 cells was observed at the concentration of 2.5 microM. The cell death rate induced by ultrasound sonification was significantly higher in the presence of hypocrellin B than in the absence of hypocrellin B. Flow cytometry showed that ultrasound exposure in the presence of hypocrellin B significantly increased the early and late apoptotic rate, 18.64% and 22.57%, respectively, compared with the controls. Nuclear condensation was observed in the nuclear staining and swollen mitochondria and more vacuolar and broken cell membrane were found in TEM after the treatment of hypocrellin B and ultrasound. Our findings demonstrated that the presence of hypocrellin B significantly enhanced the cytotoxicity of ultrasound radiation in CNE2 cells, suggesting that hypocrellin B is a novel sonosensitizer and hypocrellin B-mediated sonodynamic therapy is a potential therapeutic modality in the management of malignant tumors.

Topics: Carcinoma; Cell Death; Cell Line, Tumor; Flow Cytometry; Humans; Microscopy, Electron, Transmission; Molecular Structure; Nasopharyngeal Neoplasms; Perylene; Photosensitizing Agents; Quinones; Ultrasonic Therapy

To investigate the photodynamic inhibitory effects of Elsinochrome A (EA), Hypocrellin A (HA) and Hypocrellin B (HB) on human colorectal carcinoma Hce-8693 cells and rhesus monkey embryonic stem R366.4 cells, via inducing apoptosis.. EA, HA and HB were extracted from metabolites of Hypomyces (Fr) Tul.Sp. R366.4 cells or Hce-8693 cells were cultured with different concentrations of EA, HA or HB respectively, irradiated and incubated with fresh medium for 2 h. Cell cycle analysis was performed by flow cytometry (FCM). Data were expressed as means +/-SD and analysis of variance and Student' t-test for individual comparisons.. The photodynamic bioactivity of EA was first reported in this study. After irradiation for 5 min, 6 min, 10 min or 20 min, photoactivated EA at lower concentrations, which were 10(-7) Mol/L, 10(-6) Mol/L, 10(-5) Mol/L respectively, had no cytotoxic effects on R366.4 ES cells. Whereas, all of the three perylenequinones could induce apoptosis with a dose-dependent manner when Hce-8693 cells were incubated with photoactivated EA, HA and HB respectively. When Hce-8693 cells were incubated with EA at 10(-6) Mol/L and irradiated 5 min, 6 min, 10 min and 20 min respectively, the rates of EA-induced apoptosis were 0, 0, 13.4 % and 40.5 %. While the rates of HA-induced apoptosis were 29.5 %, 32.0 %, 40.2 % and 22.6 %. And the rates of HB-induced apoptosis were 0, 0, 0 and 13.7 % respectively. Meanwhile, after 10(-5) Mol/L treatment, the rates of EA-induced apoptosis were 32.7 %, 19.3 %, 26.4 % and 52.7 %, the rates of HA-induced apoptosis were 47.2 %, 39.1 %, 45.2 % and 56.6 %, and the rates of HB-induced apoptosis were 0, 0, 20.0 % and 13.9 % respectively.. EA, HA and HB have significant anti-cancer activity. The order of photodynamic inhibitory effects on tumor cells would be approximately HA>EA>HB. The molecular mechanisms of apoptosis may not be induced by reactive oxygen species and are worth further investigation.

Topics: Animals; Carcinoma; Cell Line; Colorectal Neoplasms; Embryo, Mammalian; Humans; Macaca mulatta; Perylene; Phenol; Photochemotherapy; Photosensitizing Agents; Quinones; Stem Cells

Novel photosensitizers Hypocrellin A (HA) and Hypocrellin B (HB), lipid soluble perylquinone derivatives of the genus Hypericum have a strong photodynamic effect on tumors and viruses. However, the mechanisms of tumor cell death induced by HA and HB are still unclear. In this study, we attempt to elucidate the photodynamic effects of HA and HB compounds in poorly differentiated (CNE2) and moderately differentiated (TW0-1) human nasopharyngeal carcinoma (NPC) cells as well as human mucosal colon (CCL-220.1) and bladder (SD) cells. Using these cell lines we investigated few hall marks of apoptotic commitments in a drug and light dose dependent manner. Tumor cells photoactivated with HA and HB showed cell size shrinkage and an increase in the sub-diploid DNA content. A loss of membrane phospholipid asymmetry associated with apoptosis was induced by all tumor cell lines as evidenced by the externalization of phosphatidylserine. Western blot analysis of poly (ADP-ribose) polymerase, a caspases substrate, showed the classical cleavage pattern (116 to 85kDa) associated with apoptosis in HA and HB-treated cell lysates. In addition, PARP cleavage was blocked by using tetrapepdide caspases inhibitors such as DEVD or z-VAD. These results demonstrate that tumor cell death induced by HB and HA is mediated by caspase proteases. This study also identifies both colon and bladder cells were more sensitive cell lines than NPC (CNE2 and TWO-1) cell lines.

Topics: Apoptosis; Carcinoma; Cell Culture Techniques; Cell Line, Tumor; Colon; Humans; Mucous Membrane; Nasopharyngeal Neoplasms; Perylene; Phenol; Photosensitizing Agents; Quinones; Urinary Bladder