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The Effect of PTEN Loss on TRPC6 Activity Involving Migration of Co-Culture Prostate Cancer & Bone Morphogenic Model

Undergraduate #11
Discipline: Biological Sciences
Subcategory: Cancer Research

Michael J. Cleveland - Tougaloo College
Co-Author(s): Collis Brown and Tamaro Hudson, Howard University, Washington, DC



Background: Prostate Cancer is the development of tumor cells in the gland in males that produces the seminal fluid for sperm. Metastatic prostate cancer is the process when cells break away from the original tumor and move into your prostate gland and begin to proliferate. In the Bone these tumors come up as bone lesions. One of the most common cell lines for prostate cancer is PC-3 and for bone cells the most common cell line is SaSo2. These cell lines are used in many research projects and have become an effective tool for elucidating cancer mechanisms. PC-3 cells have PTEN65s, which regulates cell proliferation, and the TRPC6 gene, which activates channel protein. Hypothesis: The Knockout of TPRC6 will slow down the migration of prostate cancer cells. Methods: We cultured PC-3 and SaSo2 Cells, to see the baseline for the experiment, and co-cultured together. Samples were incubated and protein was isolated. Co-culturing cells underwent transfection to knock out TRPC6. Transfected cells were cultured and protein supernatants were isolated. A Western Blot was performed to verify the knockout of the TRPC6. Cell viability assays were also conducted to test the effect of the gene loss on viability. Results: Co-cultured cells showed slow growth over time compared to the individual cell lines. Migration assays of the co-cultured cells identified that PC-3 sells traveled toward bone cells. Conclusion: The growth environment plays a role in how the cells grow and interact. The rate of cell growth can vary over the time period. The migration of the cells toward each other are affect by their environment, the Co-Cultured cells grew slower compared to the K.O. but the PC3 cells grew at the slowest rate when in 10 % serum. The morphology of the SaSo2 cells changed when TRPC6 is knocked. Significance: The overall conclusion is that TRPC6 does affect the migration of cancerous cells.

Funder Acknowledgement(s): The Office of the Provost at Howard University. Department of Veteran Affairs

Faculty Advisor: Bianca Garner, bgarner@tougaloo.edu

Role: During the researched I conducted, MTT assays, Invasion Assays, Cell Culturing,and constructing the poster and graphs.

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This material is based upon work supported by the National Science Foundation (NSF) under Grant No. DUE-1930047. Any opinions, findings, interpretations, conclusions or recommendations expressed in this material are those of its authors and do not represent the views of the AAAS Board of Directors, the Council of AAAS, AAAS’ membership or the National Science Foundation.

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