Poster Session #1: UC Ballroom
Creating a PTEN-Deficient Mesothelial Model and Growth Analysis
Presentation Type
Poster
Faculty Mentor’s Full Name
Mark A. Pershouse
Faculty Mentor’s Department
Department of Biomedical and Pharmaceutical Sciences
Abstract / Artist's Statement
Most individuals would be surprised to learn that asbestos is contained in thousands of products still marketed in the U.S. Asbestos exposures are commonplace, but few worry about the effects due to misinformation. Approximately 10% of asbestos exposures result in a deadly cancer known as malignant mesothelioma, involving lesions on the pleural and peritoneal linings of the body. Little has improved in therapy in the last 40 years. After years of negative data linking tumor suppressor PTEN to these tumors, several new publications have shown clear involvement of this gene and postulate that it is effective at encouraging tumor growth through a block to apoptosis mediated through the PTEN/PI3K/AKT/mTORc pathway. This pathway has been implicated in other cancers and is clearly one of the newest targets of cancer therapeutics.
This project aims to confirm the linkage between PTEN loss and p53 dysregulation through AKT/MDM2 in a mesothelial model system. We hypothesize that PTEN suppression by shRNA constructs will result in cells’ decreased response to apoptosis-inducing signals, as well as dysregulation of the levels of critical p53 responsive genes. The project utilizes cell lines where PTEN expression has been down-regulated to mimic tumors. In these cell lines, we hope to characterize changes in the ability of normal mesothelial cells to evade apoptosis, dysregulate cell cycle, and generally behave in a more tumor-like fashion. We isolated RNA from six cell lines and performed reverse transcription to form cDNA. In PCR, the cDNA and a Taqman probe showed varying levels of PTEN suppression. Growth curves and clone morphology showed cells’ resemblance to tumors. Research is ongoing, and our next step is developing a correlation between apoptosis and PTEN suppression levels using a Live Cell Caspase Activity Assay Kit that measures caspase 1 activation in cells after UV light exposure.
Category
Life Sciences
Creating a PTEN-Deficient Mesothelial Model and Growth Analysis
Most individuals would be surprised to learn that asbestos is contained in thousands of products still marketed in the U.S. Asbestos exposures are commonplace, but few worry about the effects due to misinformation. Approximately 10% of asbestos exposures result in a deadly cancer known as malignant mesothelioma, involving lesions on the pleural and peritoneal linings of the body. Little has improved in therapy in the last 40 years. After years of negative data linking tumor suppressor PTEN to these tumors, several new publications have shown clear involvement of this gene and postulate that it is effective at encouraging tumor growth through a block to apoptosis mediated through the PTEN/PI3K/AKT/mTORc pathway. This pathway has been implicated in other cancers and is clearly one of the newest targets of cancer therapeutics.
This project aims to confirm the linkage between PTEN loss and p53 dysregulation through AKT/MDM2 in a mesothelial model system. We hypothesize that PTEN suppression by shRNA constructs will result in cells’ decreased response to apoptosis-inducing signals, as well as dysregulation of the levels of critical p53 responsive genes. The project utilizes cell lines where PTEN expression has been down-regulated to mimic tumors. In these cell lines, we hope to characterize changes in the ability of normal mesothelial cells to evade apoptosis, dysregulate cell cycle, and generally behave in a more tumor-like fashion. We isolated RNA from six cell lines and performed reverse transcription to form cDNA. In PCR, the cDNA and a Taqman probe showed varying levels of PTEN suppression. Growth curves and clone morphology showed cells’ resemblance to tumors. Research is ongoing, and our next step is developing a correlation between apoptosis and PTEN suppression levels using a Live Cell Caspase Activity Assay Kit that measures caspase 1 activation in cells after UV light exposure.