University of Louisville
Inhibiting the Anaphase-Promoting Complex/Cyclosome: An Innovative Approach for Cancer Chemotherapy
Institution
University of Louisville
Faculty Advisor/ Mentor
J. Christopher States; Jouett Mason Hoffman; John O. Trent
Abstract
The anaphase promoting complex/cyclosome (APC/C) is a large, E3 ubiquitin ligase that regulates the cell cycle, in particular the metaphase to anaphase transition in mitosis and the reentry into G1 phase. Inhibition of the APC/C results in mitotic arrest and apoptosis in cancer cells. ANAPC2 and ANAPC11 are shown to be two vital subunits for APC/C function. in silico screening of ANAPC2 identified compounds that are predicted to prevent the association of ANAPC2 and ANAPC11. Thus, we hypothesize that the relative levels of the APC/C molecular targets, securin and cyclin B, will increase in cells treated with lead compounds. To gain better insight on the inhibition of the APC/C in cancer cells, HeLa cells were treated with lead compounds 3, 8, 10, and 11 at their respective IC50s for 24 h and then harvested to make lysates. The Bradford Protein Assay was used to determine the protein concentrations in each of the samples. To examine the relative levels of securin and cyclin B, a western blot analysis was performed. Results showed that cells treated with compounds 3, 8, 10, 11 do not have increased levels of securin and cyclin B. However, future analysis may reveal that treatment with the lead compounds causes a decrease in the levels of ubiquitinylated cyclin B and securin. This research was supported in part by University of Louisville Cancer Education Program NIH/NCI grant R25-CA134283 and a Kentucky Lung Cancer Research Program grant to JCS.
Inhibiting the Anaphase-Promoting Complex/Cyclosome: An Innovative Approach for Cancer Chemotherapy
The anaphase promoting complex/cyclosome (APC/C) is a large, E3 ubiquitin ligase that regulates the cell cycle, in particular the metaphase to anaphase transition in mitosis and the reentry into G1 phase. Inhibition of the APC/C results in mitotic arrest and apoptosis in cancer cells. ANAPC2 and ANAPC11 are shown to be two vital subunits for APC/C function. in silico screening of ANAPC2 identified compounds that are predicted to prevent the association of ANAPC2 and ANAPC11. Thus, we hypothesize that the relative levels of the APC/C molecular targets, securin and cyclin B, will increase in cells treated with lead compounds. To gain better insight on the inhibition of the APC/C in cancer cells, HeLa cells were treated with lead compounds 3, 8, 10, and 11 at their respective IC50s for 24 h and then harvested to make lysates. The Bradford Protein Assay was used to determine the protein concentrations in each of the samples. To examine the relative levels of securin and cyclin B, a western blot analysis was performed. Results showed that cells treated with compounds 3, 8, 10, 11 do not have increased levels of securin and cyclin B. However, future analysis may reveal that treatment with the lead compounds causes a decrease in the levels of ubiquitinylated cyclin B and securin. This research was supported in part by University of Louisville Cancer Education Program NIH/NCI grant R25-CA134283 and a Kentucky Lung Cancer Research Program grant to JCS.