{"id":{"repo_id":"buffalo","oai_identifier":"oai:ubir.buffalo.edu:10477/83847"},"canonical_url":"https://search.dev.ndltd.org/etd/buffalo/oai:ubir.buffalo.edu:10477/83847","repository":{"repo_id":"buffalo","name":"Buffalo","base_url":"https://ubir.buffalo.edu/oai/request"},"display":{"title":"EGF-Dependent Activation of the Oncogene AKT is Regulated by Calcium, Calmodulin and CaMKK2 in Cell Lines from Multiple Cancer Types","abstract":"M.S.","abstract_html":"M.S.","abstract_has_math":false,"creators":["Clapp, Dylan"],"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":2022,"date_issued":"2022-06-17T19:54:43Z","date_published":"2022-06-17T19:54:43Z","updated_at":"2026-07-27T19:05:28Z","subjects":["oncology","pharmaceutical sciences"],"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/10477/83847","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":["Clapp, Dylan"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2022-06-17T19:54:43Z","2020"]},{"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":["oncology","pharmaceutical sciences"]}]},{"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/10477/83847"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["M.S.","AKT is an oncogenic, serine/threonine kinase which regulates downstream targets in order to coordinate cell growth, metabolism, survival and proliferation. AKT is hyper-activated in a variety of cancer cell lines, such as OVCAR-3 (high grade, serous ovarian carcinoma). This hyperactivity is thought to promote the growth of malignant cells, and their resistance to chemotherapeutic drug treatment. The canonical pathway for the activation of AKT involves the interaction of a growth factor, like EGF, with its cognate receptor tyrosine kinase (EGFR). This initiates a cascade of reactions that result in PIP3 recruiting PDK1 and AKT to the plasma membrane where PDK1 phosphorylates AKT at (T308). Phosphorylation at (T308) is a crucial first step in AKT activation. Recent studies have shown Calcium-Calmodulin Dependent Kinase Kinase 2 (CaMKK2) to be capable of activating AKT by directly phosphorylating (T308) in multiple ovarian cancer cell lines. However, there is insufficient research on whether CaMKK2 catalyzes AKT (T308) phosphorylation in other types of cancer cells. In this thesis I examined the potential of CaMKK2 to regulate p-AKT (T308) in non-small cell lung cancer (NSCLC), epidermoid carcinoma and gastric carcinoma. The chosen cell line from each cancer type was A549, A431 and NCI-N87, respectively. I employed standard protocols to examine whether a reduction in the expression of PDK1 and CaMKK2, a reduction in intracellular calcium, and the inhibition of calmodulin affected AKT activating phosphorylation at (T308) in response to EGF. In all three cell lines, the knockdown of both PDK1 and CaMKK2 results in a decrement of p-AKT (T308) induced by EGF. The magnitude of change is very similar between the two kinases. The chelation of intracellular calcium by BAPTA-AM produces effects quantitatively similar to those seen with the knockdown of PDK1 and CaMKK2: an overall reduction in EGF-induced p-AKT (T308) levels. The inhibition of calmodulin with W7 also partially reverses the effect EGF has on raising p-AKT (T308) levels in these cells. Altogether, these findings suggest that CaMKK2, calcium and calmodulin are responsible for regulating EGF-dependent activation of AKT at (T308) in A549, A431 and NCIN87 cells.","**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":["EGF-Dependent Activation of the Oncogene AKT is Regulated by Calcium, Calmodulin and CaMKK2 in Cell Lines from Multiple Cancer Types"]}]}],"canonical_facts":{"dc:contributor":["Edelman, Arthur","Pharmacology and Toxicology"],"dc:creator":["Clapp, Dylan"],"dc:date":["2022-06-17T19:54:43Z","2020"],"dc:description":["M.S.","AKT is an oncogenic, serine/threonine kinase which regulates downstream targets in order to coordinate cell growth, metabolism, survival and proliferation. AKT is hyper-activated in a variety of cancer cell lines, such as OVCAR-3 (high grade, serous ovarian carcinoma). This hyperactivity is thought to promote the growth of malignant cells, and their resistance to chemotherapeutic drug treatment. The canonical pathway for the activation of AKT involves the interaction of a growth factor, like EGF, with its cognate receptor tyrosine kinase (EGFR). This initiates a cascade of reactions that result in PIP3 recruiting PDK1 and AKT to the plasma membrane where PDK1 phosphorylates AKT at (T308). Phosphorylation at (T308) is a crucial first step in AKT activation. Recent studies have shown Calcium-Calmodulin Dependent Kinase Kinase 2 (CaMKK2) to be capable of activating AKT by directly phosphorylating (T308) in multiple ovarian cancer cell lines. However, there is insufficient research on whether CaMKK2 catalyzes AKT (T308) phosphorylation in other types of cancer cells. In this thesis I examined the potential of CaMKK2 to regulate p-AKT (T308) in non-small cell lung cancer (NSCLC), epidermoid carcinoma and gastric carcinoma. The chosen cell line from each cancer type was A549, A431 and NCI-N87, respectively. I employed standard protocols to examine whether a reduction in the expression of PDK1 and CaMKK2, a reduction in intracellular calcium, and the inhibition of calmodulin affected AKT activating phosphorylation at (T308) in response to EGF. In all three cell lines, the knockdown of both PDK1 and CaMKK2 results in a decrement of p-AKT (T308) induced by EGF. The magnitude of change is very similar between the two kinases. The chelation of intracellular calcium by BAPTA-AM produces effects quantitatively similar to those seen with the knockdown of PDK1 and CaMKK2: an overall reduction in EGF-induced p-AKT (T308) levels. The inhibition of calmodulin with W7 also partially reverses the effect EGF has on raising p-AKT (T308) levels in these cells. Altogether, these findings suggest that CaMKK2, calcium and calmodulin are responsible for regulating EGF-dependent activation of AKT at (T308) in A549, A431 and NCIN87 cells.","**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/10477/83847"],"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":["oncology","pharmaceutical sciences"],"dc:title":["EGF-Dependent Activation of the Oncogene AKT is Regulated by Calcium, Calmodulin and CaMKK2 in Cell Lines from Multiple Cancer Types"],"dc:type":["Text","Thesis"]},"updated_at":"2026-07-27T19:05:28Z"}