Project description:Little is known about the role of miR-200s in the progression of ovarian cancer. In this study we overexpressed the miR-200c/141 cluster, the miR-200b/200a/429 cluster, or both clusters in two different murine ovarian cancer cell lines (ID8 and 28-2). As co-oeverexpression of both clusters provided the best miR-200 overexpression, we evaluated ID8 and 28-2 cells overexpressing both miR-200 clusters (ID8-200f and 28-2-200f) compared to empty vector control cells (ID8EV and 28-2EV). ID8-200f and 28-2-200f cells showed increased expression of several mesenchymal genes, decreased proliferation, decreased invasion and increased apoptosis compared to their respective controls. RNA sequencing of ID8EV, ID8-200f, 28-2EV and 28-2-200f cells revealed that overexpression of miR-200s primary regulated genes involved in epithelial mesenchymal transition EMT) and gene implicated in migration. Therefore, our work suggests that miR-200s can inhibit characteristics associated ovarian cancer progression (increased proliferation and invasion and promotion of EMT).
Project description:Ovarian cancer is the leading cause of gynecological cancer related death. The overall 5 year survival rate is only 29%. Over 85% of ovarian cancer patients present with advanced stage III or IV disease characterized by intraperitoneal metastasis when diagnosed. However, the process and mechanism of ovarian tumor metastasis remain poorly understood partially because of the lack of a mouse model which could recapitulate the development of metastatic lesion in an appropriate timeframe. In order to generate a convenient ovarian cancer model with accelerated peritoneal metastasis, we performed an in vivo selection study using ID8 ovarian cancer cells to establish a rapid metastasizing mouse ovarian cancer cell line, designated ID8-M. Syngeneic mice with intraperitoneal inoculation of ID8-M cells showed measurable ascites average 35 days after the inoculation and survived only an average of 52 days, while those inoculated with parental ID8 cells showed measurable ascites after 67 days and survived over 81 days. Further analysis showed that, compared with ID8 tumors, ID8-M tumors resulted in more macrophages in the ascites; and compared to ID8 cells, ID8-M cells were more potent to promote macrophages to acquire a M2 phenotype. A microarray analysis provided information to explain the accelerated metastatic phenotype of ID8-M cells.
Project description:Genetically modified ovarian cancer cells were used to study the role of GBP1. Proteomics-based thermal stability assay (CETSA) was performed on GBP1 knockdown and overexpressing ID8 ovarian cancer cells. Shotgun proteomics was also performed on these cells.
Project description:The data in this submission relate to whole exome sequencing from murine ovarian cancer cell line ID8. All sequencing was performed by Beckman Coulter Genomics, Grenoble, France in February 2013.
Project description:This experiment is part of a larger study examining the anti-tumour properties of interferon epsilon in ovarian cancer. The objective of this experiment was to examine the direct activity of interferon epsilon on the mouse ovarian cancer cell line, ID8, and compare it to equivalent unit concentrations of interferon beta. The goal was to determine whether interferon epsilon and interferon beta induce different patterns of gene expression in ID8 cells.
Project description:Sequencing of mRNA from ID8 tumor cells and ID8 tumor cells harvested from ascites of mice 11 weeks after intra peritoneal inoculation show acquisition of cancer stem cell-like features in ascitic tumor cells.
Project description:High grade serous ovarian cancer (HGSC) is the most common and deadly type of ovarian cancer, largely due to difficulties in early diagnosis and rapid metastasis throughout the peritoneal cavity. Previous studies have shown that expression of Notch3 correlates with worse prognosis and increased tumorigenic cell behaviors in HGSC. We investigated the mechanistic role of Notch3 in a model of metastatic ovarian cancer using the murine ovarian surface epithelial cell line, ID8 IP2. Notch3 was activated in ID8 IP2 cells via expression of the Notch3 intracellular domain (Notch3IC). Notch3IC ID8 IP2 cells injected intraperitoneally caused accelerated ascites and reduced survival compared to control ID8 IP2, particularly in early stages of disease. We interrogated downstream targets of Notch3IC in ID8 IP2 cells by RNA sequencing and found significant induction of genes that encode adhesion and extracellular matrix proteins. Notch3IC ID8 IP2 showed increased expression of ITGA1 mRNA and cell-surface protein. Notch3IC-mediated increase of ITGA1 was also seen in two human ovarian cancer cells. Notch3IC ID8 IP2 cells showed increased adhesion to collagens I and IV in vitro. We propose that Notch3 activation in ovarian cancer cells causes increased adherence to collagen-rich peritoneal surfaces. Thus, the correlation between increased Notch3 signaling and poor prognosis may be influenced by increased dissemination and metastasis of HGSC via increased attachment to the peritoneum. Implications: Expression of Notch3 accelerates the progression of ovarian cancer by increasing the adherence of disseminating cells to new metastatic sites.
Project description:This experiment is part of a larger study examining the anti-tumour properties of interferon epsilon in ovarian cancer. The goal of this experiment was to assess whether interferon signalling had been ablated in Ifnar1 CRISPR knockout (KO) ID8 cell lines, so that they could be used for in vivo models of ovarian cancer.