Project description:RNA sequencing analysis on two primary pancreatic cancer cell lines from transgenic mice: (1) cancer-collagen1 knockout cell line from KPPC;Col1pdxKO cancer-collagen1 knockout tumor and (2) control pancreatic cancer cell line from KPPC tumor
Project description:Endothelial cells (ECs) express two members of the cadherin family, VE- and N-cadherin. While VE-cadherin induces EC homotypic adhesion, N-cadherin function in ECs remains largely unknown. EC-specific inactivation of either VE- or N-cadherin leads to early foetal lethality suggesting that these cadherins play a non-redundant role in vascular development. Goal of this study was to further investigate this hypothesis analyzing both additive and divergent functions of the two cadherins in ECs. The three endothelial cell lines were cultured. Total RNA was extracted using commercial homogenization (QIAshredder) and purification (RNeasy Mini Kit) reagents (Qiagen). Quality control (QC) of the RNA samples was performed using an Agilent Bioanalyzer 2100 (Agilent Technologies). Two different RNA extractions were processed for each of the cell lines under analysis, and each sample was labelled and hybridized to a Mouse Gene 1.0 ST Genechip array according to the manufacturer’s specifications (Affymetrix Inc). Data were analysed using Partek Genomics Suite v6.3 software (RMA algorithm). Differentially expressed genes were identified through ANOVA, using a fold change cutoff >2 and a p-value of 0.05.
Project description:Endothelial cells (ECs) express two members of the cadherin family, VE- and N-cadherin. While VE-cadherin induces EC homotypic adhesion, N-cadherin function in ECs remains largely unknown. EC-specific inactivation of either VE- or N-cadherin leads to early foetal lethality suggesting that these cadherins play a non-redundant role in vascular development. Goal of this study was to further investigate this hypothesis analyzing both additive and divergent functions of the two cadherins in ECs.
Project description:The aim of the experiment was to compare a newly defined population VE-Cadherin+GFP+ to control populations, VE-Cadherin- GFP+ and VE-Cadherin+GFP-.
Project description:Gene expression was compared between E18.5 E-cadherin conditional knockout (cKO) small intestine and E18.5 control mouse small intestine. E18.5 mouse small intestine was collected from control and E-cadherin conditional knockout mice. RNA was prepared. Affymetrix Mouse Gene 1.0 gene arrays were used to interrogate gene expression. Data was analyzed using dChip software.
Project description:Gene expression was compared between E18.5 E-cadherin conditional knockout (cKO) small intestine and E18.5 control mouse small intestine.
Project description:Interventions: experimental group :PD-1 Knockout Engineered T Cells
Primary outcome(s): Number of participants with Adverse Events and/or Dose Limiting Toxicities as a Measure of Safety and tolerability of dose of PD-1 Knockout T cells using Common Terminology Criteria for Adverse Events (CTCAE v4.0) in patients
Study Design: historical control
Project description:Diffuse-type gastric cancer (DGC) exhibits rapid disease progression and a poor patient prognosis. We have previously established an E-cadherin/p53 double conditional knockout (DCKO) mouse line as the first genetically engineered one, which morphologically and molecularly recapitulates human DGC. In this study, we explored low-molecular-weight drugs selectively eliminating mouse DGC cells, and then validated their inhibitory effects on human DGC. We first derived mouse gastric cancer (GC) cell lines from DGC of the DCKO mice, which notably demonstrated enhanced tumorigenic activity in immunodeficient mice and acquired tolerance to cytotoxic anti-cancer agents. We next performed a synthetic lethal screening of 1535 annotated chemical compounds by using them. Comparing cell viability of the E-cadherin/p53-deficient GC and p53-deficient gastric epithelial (GE) cells under treatment with the compound library, we identified 27 candidates with specific toxicity to the GC cell lines. The most potent drug mestranol, an estrogen derivative, and other estrogen receptor modulators induced apoptotic events preceded by DNA damage only in the GC cell lines, but not in the GE. Moreover, mestranol could significantly suppress tumor growth of the GC cells subcutaneously transplanted into nude mice, consistent with longer survival time in the female DCKO mice than in the male. As expected, human E-cadherin-mutant and -low gastric cancer cells showed higher susceptibility to estrogen drugs in contrast to E-cadherin-intact ones in vitro and in vivo. These findings may lead to the development of novel therapeutic strategies targeting E-cadherin-deficient DGC.