{"id":{"repo_id":"buffalo","oai_identifier":"oai:ubir.buffalo.edu:10477/86805"},"canonical_url":"https://search.dev.ndltd.org/etd/buffalo/oai:ubir.buffalo.edu:10477/86805","repository":{"repo_id":"buffalo","name":"Buffalo","base_url":"https://ubir.buffalo.edu/oai/request"},"display":{"title":"Genotoxic Stress Causes Activation of Akt via Ca2+/Calmodulin-Dependent Protein Kinase Kinase 2 Pathway","abstract":"M.S.","abstract_html":"M.S.","abstract_has_math":false,"creators":["Shetty, Sanjana; 0000-0002-2707-1349"],"institution":"State University of New York at Buffalo","degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":["Edelman, Arthur","Pharmacology and Toxicology"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2025,"date_issued":"2025-02-25T23:22:56Z","date_published":"2025-02-25T23:22:56Z","updated_at":"2026-07-27T19:05:37Z","subjects":["pharmacology"],"languages":["eng"],"rights":["Users of works found in University at Buffalo Institutional Repository (UBIR) are responsible for identifying and contacting the copyright owner for permission to reuse. University at Buffalo Libraries do not manage rights for copyright-protected works and cannot assist with permissions.","Copyright retained by author."],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"http://hdl.handle.net/10477/86805","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Edelman, Arthur","Pharmacology and Toxicology"]},{"key":"dc:creator","label":"Author","values":["Shetty, Sanjana; 0000-0002-2707-1349"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2025-02-25T23:22:56Z","2020","2020-07-11 19:26:36"]},{"key":"dc:publisher","label":"Institution","values":["State University of New York at Buffalo"]},{"key":"dc:type","label":"Dc Type","values":["Text","Thesis"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["pharmacology"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["eng"]},{"key":"dc:rights","label":"Dc Rights","values":["Users of works found in University at Buffalo Institutional Repository (UBIR) are responsible for identifying and contacting the copyright owner for permission to reuse. University at Buffalo Libraries do not manage rights for copyright-protected works and cannot assist with permissions.","Copyright retained by author."]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["http://hdl.handle.net/10477/86805"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["M.S.","One of the major obstacles to the successful treatment of ovarian cancer is the rapid development of resistance to chemotherapy. Many genotoxic cancer treatments activate the serine/threonine kinase (Akt) pathway. Activation of this pathway plays a very important role in essential cellular functions such as survival, proliferation, migration, and angiogenesis that underlie the biology of human cancer. However, the mechanism by which Akt is activated in response to DNA damaging chemotherapeutic drugs and its effects on downstream targets are still unclear. In this study, we find that the treatment of ovarian cancer cells with the chemotherapeutic topoisomerase II inhibitor etoposide results in activating phosphorylation of Akt. A previous study from our laboratory reported that although the PI3K/ phosphoinositide-dependent kinase 1 (PDK1) pathway represents the canonical mechanism for phosphorylation of Akt at Thr-308, a non-canonical mechanism involves mediation of Akt phosphorylation at Thr-308 via Ca2+ /Calmodulin (CaM)-dependent protein kinase kinase 2 (CaMKK2). Since CaMKK2 is highly expressed in high-grade serous ovarian cancer, we investigated its role in Akt activation in ovarian cancer cells response to genotoxic stress caused by etoposide. We observed that Akt phosphorylation was significantly enhanced by etoposide in OVCAR-3 ovarian cancer cells and that RNAi-mediated knockdown of CaMKK2 significantly reduced the increase in activating Akt phosphorylation caused by etoposide. Consistent with an effect on p-Akt Thr-308 was an observed reduction in phosphorylation of the Akt downstream target PRAS40. Treatment of OVCAR3 cells with the intracellular calcium chelator, BAPTA-AM, or inhibition with the calmodulin inhibitor W-7 also decreased phosphorylation of Akt verifying that the action of CaMKK2 is reliant on its Ca2+/CaM-dependence.","**To request an accessible version of the file(s) associated with this item, contact library@buffalo.edu. Please include the item's persistent URL [http://hdl.handle.net/. . .] in your request.**"]},{"key":"dc:format","label":"Dc Format","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Genotoxic Stress Causes Activation of Akt via Ca2+/Calmodulin-Dependent Protein Kinase Kinase 2 Pathway"]}]}],"canonical_facts":{"dc:contributor":["Edelman, Arthur","Pharmacology and Toxicology"],"dc:creator":["Shetty, Sanjana; 0000-0002-2707-1349"],"dc:date":["2025-02-25T23:22:56Z","2020","2020-07-11 19:26:36"],"dc:description":["M.S.","One of the major obstacles to the successful treatment of ovarian cancer is the rapid development of resistance to chemotherapy. Many genotoxic cancer treatments activate the serine/threonine kinase (Akt) pathway. Activation of this pathway plays a very important role in essential cellular functions such as survival, proliferation, migration, and angiogenesis that underlie the biology of human cancer. However, the mechanism by which Akt is activated in response to DNA damaging chemotherapeutic drugs and its effects on downstream targets are still unclear. In this study, we find that the treatment of ovarian cancer cells with the chemotherapeutic topoisomerase II inhibitor etoposide results in activating phosphorylation of Akt. A previous study from our laboratory reported that although the PI3K/ phosphoinositide-dependent kinase 1 (PDK1) pathway represents the canonical mechanism for phosphorylation of Akt at Thr-308, a non-canonical mechanism involves mediation of Akt phosphorylation at Thr-308 via Ca2+ /Calmodulin (CaM)-dependent protein kinase kinase 2 (CaMKK2). Since CaMKK2 is highly expressed in high-grade serous ovarian cancer, we investigated its role in Akt activation in ovarian cancer cells response to genotoxic stress caused by etoposide. We observed that Akt phosphorylation was significantly enhanced by etoposide in OVCAR-3 ovarian cancer cells and that RNAi-mediated knockdown of CaMKK2 significantly reduced the increase in activating Akt phosphorylation caused by etoposide. Consistent with an effect on p-Akt Thr-308 was an observed reduction in phosphorylation of the Akt downstream target PRAS40. Treatment of OVCAR3 cells with the intracellular calcium chelator, BAPTA-AM, or inhibition with the calmodulin inhibitor W-7 also decreased phosphorylation of Akt verifying that the action of CaMKK2 is reliant on its Ca2+/CaM-dependence.","**To request an accessible version of the file(s) associated with this item, contact library@buffalo.edu. Please include the item's persistent URL [http://hdl.handle.net/. . .] in your request.**"],"dc:format":["application/pdf"],"dc:identifier":["http://hdl.handle.net/10477/86805"],"dc:language":["eng"],"dc:publisher":["State University of New York at Buffalo"],"dc:rights":["Users of works found in University at Buffalo Institutional Repository (UBIR) are responsible for identifying and contacting the copyright owner for permission to reuse. University at Buffalo Libraries do not manage rights for copyright-protected works and cannot assist with permissions.","Copyright retained by author."],"dc:subject":["pharmacology"],"dc:title":["Genotoxic Stress Causes Activation of Akt via Ca2+/Calmodulin-Dependent Protein Kinase Kinase 2 Pathway"],"dc:type":["Text","Thesis"]},"updated_at":"2026-07-27T19:05:37Z"}