| Background and objective:Ovarian cancer is a serious threat to life and health of women. The morbidity of the patients with ovarian cancer reaches about47%, which is ranking the first in gynecological malignancies.Due to hidden symptomps of the patients at early stage and lack of effective detection means, more than70%of the patients were found at advanced stage, The current therapies have many shortcomings with limited success. Photodynamic therapy is capable to selectively kill cancer cells and has many advantages over the current therapies such as radiotherapy and chemotherapy. Hypocrellin B is a naturally occurred product from a traditional Chinese herb, which mainly distributes in yunnan province China. It has role of anti-inflammatory, antibacteria, analgesia, local anesthesia. Recent studies have found that hypocrellin B is an excellent photo sens itizer for photodynamic therapy This project arms to investigate the effect of hypocrellin B-mediated photodynamic therapy on apoptosis, intends to explore the killing effect and the destruction mechanism of hyporellin B-induced photodynamic therapy on ovarian cancer HO8910cells and observes the impact on the metastatic ability of the cells.Materials/Methods:The HO8910cells were incubated with hyporellin B concentration at2.5μM for5h in the dark and then were irradiated by LED light source with the wavelength of470nm and the power density of60mW/cm2. photo cytotoxicity of hypocrellin B-induced photodynamic therapy on HO-8910cells was assessed using the3-(4,5-dimthylthiazol-2-yl)-2,5diphenyl-tetrazolium bromide (MTT) reduction assay and the morphological changes of the HO-8910cells by light microscope. Apoptosis of the HO-8910cells was measure using flow cytometer (FCM) with Annexin V-FITC/PI staining and the nuclear staining with Hoechst33258.Ultrastructure was observed using transmission electron microscopy (TEM). Mitochondrial membrane potential was evaluated using FCM analysis with JC-1staining. Intracellular reaction oxygen species were detected by DCFH-DA staining. Finally, clone assay, invasion assay, migration assay, and adhesion assay were investigated in this study.Results:The photo cytotoxicity of hyporellin B-induced photodynamic therapy on HO-8910cells was the dramatically significant, and light dose-dependent photo cytotoxicity in the HO-8910cells was observed. No significant cytotoxicity was observed in other groups; Light morphology showed that the number of HO-8910cells after PDT treatment was significantly reduced. Nuclear staining found that nuclear condensation and typical apoptotic body in PDT treated cells. FCM with Annexin V-FITC/PI staining showed the apoptotic rate of PDT treatment group after light-activated6h was significantly higher than other groups, and the apoptosis bodies were also found in TEM. The collapse of mitochondrial membrane potential and the increase of ROS in PDT treatment group after light-activated3h were more obvious than other groups. The adhesive, migrative and invasive ability of ovarian cancer HO-8910cells were significant inhibited after hypocrellin B-induced photodynamic therapy with light dose0.4J/cm2.Conclusion:Hypocrellin B-induced photodynamic significantly damaged ovarian cancer cells and ROS increase was probably an important cause inducing apoptosis. Moreover, hypocrellin B-madiated PDT inhibited adhesion, migration and invasion of ovarian cancer cells, which suggests hypocrellin B-mediated PDT is probably beneficial to the inhibition of ovarian cancer spread. Therefore, photodynamic therapy with hypocrellin B show a promising in the management of ovarian cancer. |
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