Apoptotic And Autophagic Cell Death In Ganoderic Acid Dm Treated Prostate Cancer Cells

Nathan Leaphart

School Name

South Carolina Governor's School for Science and Mathematics

Grade Level

12th Grade

Presentation Topic

Microbiology

Presentation Type

Mentored

Mentor

Mentor: Azizul Haque, Department of Microbiology and Immunology, Medical University of South Carolina

Abstract

Traditional treatments of prostate cancer have difficulty handling the metastatic form of the cancer. Immense research has been done on discovering a more “user-friendly” chemoimmunotherapeutic. One potential chemoimmunotherapeutic is Ganoderic Acid DM (GA-DM). To understand the effects of GA-DM on advanced prostate cancer, two human prostate cancer cell lines, LnCaP and PC-3, were used in running and analyzing MTS assays, Western blots, and Annexin V and PI staining tests. To understand autophagy in GA-DM treated prostate cancer cells, a MTS assay was ran with the autophagy inhibitor 3-MA. Results showed that GA-DM induced differential autophagy in the two different prostate cancer cell lines. With autophagy inhibited, GA- DM treated LnCaP expressed a decrease in cytotoxicity, while GA-DM treated PC-3 expressed an increase in cytotoxicity. To analyze Bax protein expression in the cells, a Western blot was conducted. This Western blot proved that Bax expression did not changed between GA-DM treated and untreated LnCaP; however, the Bax expression for GA-DM treated and untreated PC-3 was unable to be interpreted. To quantify the amount of apoptotic cell death occurring in GA-DM treated PC-3, an Annexin V and PI staining test was organized. The staining showed that a 40 μM GA-DM treatment increased the amount of cells in early apoptosis by 16.75% and the amount of cells in late apoptosis by 56.03%. Conclusively, these experiments showed that differential autophagy occurs in GA-DM treated prostate cancer cells, and that GA-DM induces apoptosis in prostate cancer cells.

Start Date

4-11-2015 8:45 AM

End Date

4-11-2015 9:00 AM

COinS
 
Apr 11th, 8:45 AM Apr 11th, 9:00 AM

Apoptotic And Autophagic Cell Death In Ganoderic Acid Dm Treated Prostate Cancer Cells

Traditional treatments of prostate cancer have difficulty handling the metastatic form of the cancer. Immense research has been done on discovering a more “user-friendly” chemoimmunotherapeutic. One potential chemoimmunotherapeutic is Ganoderic Acid DM (GA-DM). To understand the effects of GA-DM on advanced prostate cancer, two human prostate cancer cell lines, LnCaP and PC-3, were used in running and analyzing MTS assays, Western blots, and Annexin V and PI staining tests. To understand autophagy in GA-DM treated prostate cancer cells, a MTS assay was ran with the autophagy inhibitor 3-MA. Results showed that GA-DM induced differential autophagy in the two different prostate cancer cell lines. With autophagy inhibited, GA- DM treated LnCaP expressed a decrease in cytotoxicity, while GA-DM treated PC-3 expressed an increase in cytotoxicity. To analyze Bax protein expression in the cells, a Western blot was conducted. This Western blot proved that Bax expression did not changed between GA-DM treated and untreated LnCaP; however, the Bax expression for GA-DM treated and untreated PC-3 was unable to be interpreted. To quantify the amount of apoptotic cell death occurring in GA-DM treated PC-3, an Annexin V and PI staining test was organized. The staining showed that a 40 μM GA-DM treatment increased the amount of cells in early apoptosis by 16.75% and the amount of cells in late apoptosis by 56.03%. Conclusively, these experiments showed that differential autophagy occurs in GA-DM treated prostate cancer cells, and that GA-DM induces apoptosis in prostate cancer cells.