AKT inhibitors promote cell death in cervical cancer through disruption of mTOR signaling and glucose uptake.
PI3K/AKT pathway alterations are associated with incomplete response to chemoradiation in human cervical cancer. This study was performed to test for mutations in the PI3K pathway and to evaluate the effects of AKT inhibitors on glucose uptake and cell viability.



Mutational analysis of DNA from 140 pretreatment tumor biopsies and 8 human cervical cancer cell lines was performed. C33A cells (PIK3CAR88Q and PTENR233*) were treated with increasing concentrations of two allosteric AKT inhibitors (SC-66 and MK-2206) with or without the glucose analogue 2-deoxyglucose (2-DG). Cell viability and activation status of the AKT/mTOR pathway were determined in response to the treatment. Glucose uptake was evaluated by incubation with 18F-fluorodeoxyglucose (FDG). Cell migration was assessed by scratch assay.



Activating PIK3CA (E545K, E542K) and inactivating PTEN (R233*) mutations were identified in human cervical cancer. SC-66 effectively inhibited AKT, mTOR and mTOR substrates in C33A cells. SC-66 inhibited glucose uptake via reduced delivery of Glut1 and Glut4 to the cell membrane. SC-66 (1 µg/ml-56%) and MK-2206 (30 µM-49%) treatment decreased cell viability through a non-apoptotic mechanism. Decreases in cell viability were enhanced when AKT inhibitors were combined with 2-DG. The scratch assay showed a substantial reduction in cell migration upon SC-66 treatment.



The mutational spectrum of the PI3K/AKT pathway in cervical cancer is complex. AKT inhibitors effectively block mTORC1/2, decrease glucose uptake, glycolysis, and decrease cell viability in vitro. These results suggest that AKT inhibitors may improve response to chemoradiation in cervical cancer.
Department of Radiation Oncology, Washington University School of Medicine, St. Louis, Missouri, United States of America.
Ramachandran
R
Rashmi
Department of Radiation Oncology, Washington University School of Medicine, St. Louis, Missouri, United States of America.
Carl
C
DeSelm
Department of Genetics, Washington University School of Medicine, St. Louis, Missouri, United States of America.
Cynthia
C
Helms
Department of Genetics, Washington University School of Medicine, St. Louis, Missouri, United States of America; Division of Gynecologic Oncology, Washington University School of Medicine, St. Louis, Missouri, United States of America.
Anne
A
Bowcock
Department of Radiation Oncology, Washington University School of Medicine, St. Louis, Missouri, United States of America; Alvin J. Siteman Cancer Center, Washington University School of Medicine, St. Louis, Missouri, United States of America.
Buck E
BE
Rogers
Janet L
JL
Rader
Dept of Obstetrics and Gynecology, Medical College of Wisconsin, Milwaukee, Wisconsin, United States of America.
Janet
J
Rader
Department of Radiation Oncology, Washington University School of Medicine, St. Louis, Missouri, United States of America; Division of Nuclear Medicine, Mallinckrodt Institute of Radiology, Washington University School of Medicine, St. Louis, Missouri, United States of America; Division of Gynecologic Oncology, Washington University School of Medicine, St. Louis, Missouri, United States of America; Alvin J. Siteman Cancer Center, Washington University School of Medicine, St. Louis, Missouri, United States of America.
Perry W
PW
Grigsby
Department of Radiation Oncology, Washington University School of Medicine, St. Louis, Missouri, United States of America; Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, Missouri, United States of America; Alvin J. Siteman Cancer Center, Washington University School of Medicine, St. Louis, Missouri, United States of America.
Julie K
JK
Schwarz
D000970:Antineoplastic Agents / Q000494:pharmacology; D016923:Cell Death / Q000187:drug effects; D045744:Cell Line, Tumor; D058534:Class I Phosphatidylinositol 3-Kinases; D003512:Cyclohexanones / Q000494:pharmacology*; D005260:Female; D005947:Glucose / Q000378:metabolism*; D006575:Heterocyclic Compounds, 3-Ring / Q000494:pharmacology*; D006801:Humans; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D000081082:Phosphoinositide-3 Kinase Inhibitors; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D051057:Proto-Oncogene Proteins c-akt / Q000037:antagonists & inhibitors*; D011725:Pyridines / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects; D058570:TOR Serine-Threonine Kinases / Q000378:metabolism*; D002583:Uterine Cervical Neoplasms / Q000378:metabolism / Q000473:pathology*
D000970:Antineoplastic Agents; D058534:Class I Phosphatidylinositol 3-Kinases; D003512:Cyclohexanones; D005947:Glucose; D006575:Heterocyclic Compounds, 3-Ring; D019869:Phosphatidylinositol 3-Kinases; D000081082:Phosphoinositide-3 Kinase Inhibitors; D047428:Protein Kinase Inhibitors; D051057:Proto-Oncogene Proteins c-akt; D011725:Pyridines; D058570:TOR Serine-Threonine Kinases; C548887:MK 2206; C000598518:SC-66 compound; C546842:MTOR protein, human; C484760:PIK3CA protein, human