Title

Expression Of St2 Soluble Receptor In Mc38 And Ct26 Colon Cancer Cells

Author(s)

Harper English

School Name

Governor's School for Science and Math

Grade Level

12th Grade

Presentation Topic

Cell and Molecular Biology

Presentation Type

Mentored

Mentor

Mentor: Dr. Marjorette Peña; Department of Biology, University of South Carolina

Abstract

Colorectal cancer is the second leading cause of cancer related deaths in the United States, accounting for almost 10% of the deaths associated with cancer. The ultimate cause of death is typically liver metastasis. Survival rates drop from 90% to less than 10% within the first five years of metastasis formation. Metastasis results from signaling caused by the binding of receptors on the cancer cell. The signaling of receptors like IL1R1/ST2 in cancer cells will lead to mast cell degranulation. Interleukin-33 (IL-33), a cytokine, binds to IL1R1/ST2 receptors on the cancer cell. There are two variants of the ST2 receptor, membrane embedded ST2L and soluble sST2. If IL-33 binds to ST2L on the cell membrane, pathways are activated speeding up metastasis formation. If IL-33 binds to the soluble form, sST2 acts as a decoy receptor preventing signaling across the cell membrane. Increasing sST2 levels in CT26 and MC38 colon cancer cells may decrease IL-33 signaling and reduce the probability of metastasis. We constructed an overexpression plasmid containing sST2 cDNA under the control of a cytomegalovirus (CMV) promotor. We verified the plasmid construction by gel electrophoresis. The sST2 overexpression plasmid was transformed into the CT26 and MC38 cancer cells. SST2 transcription increased approximately 10-12 fold in transformed cells, as determined by RealTime PCR. We verified overexpression of the sST2 protein by Western blot. With a plasmid that now generates an overexpression of sST2, it can now be tested on mice to determine if the chances of metastasis formation are lowered.

Location

Owens 201

Start Date

4-16-2016 10:00 AM

COinS
 
Apr 16th, 10:00 AM

Expression Of St2 Soluble Receptor In Mc38 And Ct26 Colon Cancer Cells

Owens 201

Colorectal cancer is the second leading cause of cancer related deaths in the United States, accounting for almost 10% of the deaths associated with cancer. The ultimate cause of death is typically liver metastasis. Survival rates drop from 90% to less than 10% within the first five years of metastasis formation. Metastasis results from signaling caused by the binding of receptors on the cancer cell. The signaling of receptors like IL1R1/ST2 in cancer cells will lead to mast cell degranulation. Interleukin-33 (IL-33), a cytokine, binds to IL1R1/ST2 receptors on the cancer cell. There are two variants of the ST2 receptor, membrane embedded ST2L and soluble sST2. If IL-33 binds to ST2L on the cell membrane, pathways are activated speeding up metastasis formation. If IL-33 binds to the soluble form, sST2 acts as a decoy receptor preventing signaling across the cell membrane. Increasing sST2 levels in CT26 and MC38 colon cancer cells may decrease IL-33 signaling and reduce the probability of metastasis. We constructed an overexpression plasmid containing sST2 cDNA under the control of a cytomegalovirus (CMV) promotor. We verified the plasmid construction by gel electrophoresis. The sST2 overexpression plasmid was transformed into the CT26 and MC38 cancer cells. SST2 transcription increased approximately 10-12 fold in transformed cells, as determined by RealTime PCR. We verified overexpression of the sST2 protein by Western blot. With a plasmid that now generates an overexpression of sST2, it can now be tested on mice to determine if the chances of metastasis formation are lowered.