Title

Studying the effects of PMA induced differentiation on cell signaling on actin cytoskeleton proteins in THP-1 cells

Submitter Information

Shyane R. WiegersFollow

Streaming Media

Type of Proposal

Oral presentation

Start Date

29-3-2016 8:30 AM

End Date

29-3-2016 9:50 AM

Faculty

Faculty of Science

Faculty Sponsor

Dr. Andrew Hubberstey

Lancer's Lair

1

Abstract

The mechanism of cell movement is complex and has much to be explored. Previous literature has discovered that the actin cytoskeleton is the major source of internally created force that regulates cell shape and helps in migration (Pollard & Cooper, 2009). A common model system that has been used to study underlying molecular mechanisms in monocytes and macrophages are a monocytic leukemia suspension cell line known as THP-1 cells. The movement required for macrophages to travel around the body requires interactions from several cytoskeleton proteins, and should thus have different levels than premature monocytes. Assessing the levels of protein and comparing them at different time points in the differentiation process should give a greater insight of the proteins that regulate actin cytoskeleton and affect cell movement. To gain a more comprehensive understanding of how cell signaling effects cytoskeleton proteins such as cofilin, WDR-1 (WDR repeat protein) and CAPs (Cyclase Associated Protein), THP-1 cells will be exposed to Phorbol 12-myristate 13-acetate (PMA) to induce cell differentiation from premature monocytes into macrophages. Cells will then be lysed at different time points, and subjected to western blot analysis to analyze the levels of such cytoskeleton proteins. The investigation of cell signaling in actin cytoskeleton proteins in THP-1 cells may provide insight into drug therapy treatments for preventing the movement of cancerous cells.

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Mar 29th, 8:30 AM Mar 29th, 9:50 AM

Studying the effects of PMA induced differentiation on cell signaling on actin cytoskeleton proteins in THP-1 cells

The mechanism of cell movement is complex and has much to be explored. Previous literature has discovered that the actin cytoskeleton is the major source of internally created force that regulates cell shape and helps in migration (Pollard & Cooper, 2009). A common model system that has been used to study underlying molecular mechanisms in monocytes and macrophages are a monocytic leukemia suspension cell line known as THP-1 cells. The movement required for macrophages to travel around the body requires interactions from several cytoskeleton proteins, and should thus have different levels than premature monocytes. Assessing the levels of protein and comparing them at different time points in the differentiation process should give a greater insight of the proteins that regulate actin cytoskeleton and affect cell movement. To gain a more comprehensive understanding of how cell signaling effects cytoskeleton proteins such as cofilin, WDR-1 (WDR repeat protein) and CAPs (Cyclase Associated Protein), THP-1 cells will be exposed to Phorbol 12-myristate 13-acetate (PMA) to induce cell differentiation from premature monocytes into macrophages. Cells will then be lysed at different time points, and subjected to western blot analysis to analyze the levels of such cytoskeleton proteins. The investigation of cell signaling in actin cytoskeleton proteins in THP-1 cells may provide insight into drug therapy treatments for preventing the movement of cancerous cells.