| Background and ObjectiveThe mortality of ovarian cancer is the highest in female genital system malignant tumors. Because of its insidious onset, lack of specific symptoms and effective diagnostic and therapeutic tools result in advanced stage and poor prognosis when diagnosed. At present, the treatment of ovarian cancer is surgical excision supplemented by radiotherapy, chemotherapy and traditional Chinese medicine, but the effect is not outstanding. In recent years, the research on tumor has gradually expanded to the cellular and molecular level, and ovarian cancer is no exception. Sigal transduction pathways between cells and mechanism of the tumorigenesis and progression are research hotspots for years.Hedgehog( Hh) signaling pathway has been firstly discovered in the regulation of embryonic segments development in Drosophila. Later, the biochemical and functional homologs of Drosophila Hh signaling genes have also been isolated in vertebrates( includinghuman). Hh signaling pathways as one of the classic signaling pathways in the body, there is a close relationship between not only Hh signaling pathways and embryonic development, tissue turnover, but also between Hh signaling pathways and the initiation and progression of human tumors. At present, the study about the role of Hh signaling pathway in gynecological tumors such as cervical cancer, endometrial cancer, etc has formed rather mature method. Abnormal activation of Hh signaling pathway exists in ovarian cancer tissues obviously. While inhibition of this pathway will lead to inhibition of ovarian cancer growth and apoptosis of cancer cells. Studies have indicated that the abnormal activation of Hh signaling plays an important role in the occurrence and development of ovarian cancer. Therefore, the inhibitors of Hh signaling molecules may be one of the ways for ovarian cancer.GLI proteins, as a member of Hh signaling pathways downstream, mediating most Hh signaling pathways, have abnormal activation and proliferation in a variety of common cancers, such as breast cancer and glioma. It has been found that such as GANT61, GANT58 and HPI series are downstream small molecule inhibitors, which inhibit the proliferation of tumor cell through the inhibition of GLI expression inhibition region. The study of Hh signaling pathway in ovarian cancer is more concentrated in the upper reaches of the site, but there is less research on the downstream sites. But so far, reports about HPI-4 in ovarian cancer are rarely in the world and the mechanism is still not very clear.Cell cycle regulation disorder and dysregulation exist in almost all tumor, and it could be argued that tumor is a disaese of disorder of cell cycle modulation. CyclinD1, as extremely important regulation proteins of the nucleus in the cell cycle, its excessive expression can lead to the G1/S defect and can shorten the G1 period.The cells overpass G1/S restriction point and loss its anti-proliferation function, finally turned into abnormal cells. The abnormal cells out of the regulation of cell cycle, continuously proliferate and cause carcinogenesis. Abnormal expression of cyclinD1 protein plays an important role in the uncontrol of cell cycle, abnormal cellular proliferation and cancerization.This study is to observe the effect of HPI-4, a Hh singaling pathway inhibitor, on the proliferation, cell cycle distribution and apoptosis of human epithelial ovarian cancer A2780 cells and on the expression of CyclinD1 and GLI1. In order to further explore probable mechanisms of Hh signaling pathways involved in the development and occurrence of ovarian cancer. This study provides a new target for the clinical therapy of ovarian cancer and lay a solid theoretical foundation for the further study of the small molecule inhibitor HPI-4 in ovarian cancer. Materials and MethodsBy culturing in vitro and recovery of human epithelial ovarian cancer A2780 cells to the exponential phase of growth, firstly added to the different concentrations of Hh singaling pathway inhibitor HPI-4 to A2780 cells of post-exponential phase and observed the apoptosis rate of A2780 cells by MTT assay; then the changes of cell cycle and apoptosis were detected by flow cytometry; finally western blot to evaluate the expression of CyclinD1 and GLI1 and in human epithelial ovarian cancer A2780 cells in vitro treated by different concentrations of HPI-4 acting 48 hours.SPSS20.0 Statistical analysis software was used to analysis related experiental data ?=0.05 was considered as the detection standard. Results1?MTT assay showed with the increasing of HPI-4 concentration and with the extension of drug action time, the apoptosis of A2780 cells actually gradually increased, exhibiting concentration- and time-dependdent manners(P<0.05).2?In analyses of A2780 cells by flow cytometry, with the increasing of HPI-4, the percentage of cells in G0~G1 phase also increased gradually, and cells in S decreased gradually. Cells that entered the DNA synthesis phase were gradually reduced. The apoptosis was gradually increased, and there was a significant difference(P<0.05). G2 /M phase cells in the fluorescent signal expressed no significant difference.3?According to the western blot test analysis,we examined the upregulated expressions of CyclinD1 and GLI1. At the same time, it revealed that CyclinD1 and GLI1 significantly declined under the influence of HPI-4, and showed that the expression level of CyclinD1 and GLI1 in A2780 cells reduced with the increase of HPI-4. Conclusions1?HPI-4, as a Hh singaling pathway inhibitor, can effectively inhibit the proliferation of human epithelial ovarian cancer A2780 cells, exhibiting concentration and time dependent manners.2?HPI-4 can prevent from synthesizing DNA and induce apoptosis of human epithelial ovarian cancer A2780 cells.3?Abnormal expression of CyclinD1 and GLI1 protein in human epithelial ovarian cancer A2780 cells associated with the activation of Hh signaling pathway. |
PDF Full Text Request