title: Lethality of PAK3 and SGK2 shRNAs to human papillomavirus positive cervical cancer cells is independent of PAK3 and SGK2 knockdown.
abstract: The p21-activated kinase 3 (PAK3) and the serum and glucocorticoid-induced kinase 2 (SGK2) have been previously proposed as essential kinases for human papillomavirus positive (HPV+) cervical cancer cell survival. This was established using a shRNA knockdown approach. To validate PAK3 and SGK2 as potential targets for HPV+ cervical cancer therapy, the relationship between shRNA-induced phenotypes in HPV+ cervical cancer cells and PAK3 or SGK2 knockdown was carefully examined. We observed that the phenotypes of HPV+ cervical cancer cells induced by various PAK3 and SGK2 shRNAs could not be rescued by complement expression of respective cDNA constructs. A knockdown-deficient PAK3 shRNA with a single mismatch was sufficient to inhibit HeLa cell growth to a similar extent as wild-type PAK3 shRNA. The HPV+ cervical cancer cells were also susceptible to several non-human target shRNAs. The discrepancy between PAK3 and SGK2 shRNA-induced apoptosis and gene expression knockdown, as well as cell death stimulation, suggested that these shRNAs killed HeLa cells through different pathways that may not be target-specific. These data demonstrated that HPV+ cervical cancer cell death was not associated with RNAi-induced PAK3 and SGK2 knockdown but likely through off-target effects.
authors.affiliation: Department of Virology, R&D of Bristol-Myers Squibb Company, Wallingford, Connecticut, United States of America.
authors.forename: Nannan
authors.initials: N
authors.lastname: Zhou
authors.affiliation: Department of Virology, R&D of Bristol-Myers Squibb Company, Wallingford, Connecticut, United States of America.
authors.forename: Bo
authors.initials: B
authors.lastname: Ding
authors.affiliation: Department of Leads Discovery, R&D of Bristol-Myers Squibb Company, Wallingford, Connecticut, United States of America.
authors.forename: Michele
authors.initials: M
authors.lastname: Agler
authors.affiliation: Department of Virology, R&D of Bristol-Myers Squibb Company, Wallingford, Connecticut, United States of America.
authors.forename: Mark
authors.initials: M
authors.lastname: Cockett
authors.affiliation: Department of Virology, R&D of Bristol-Myers Squibb Company, Wallingford, Connecticut, United States of America.
authors.forename: Fiona
authors.initials: F
authors.lastname: McPhee
mesh_terms: D017209:Apoptosis; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D005260:Female; D055785:Gene Knockdown Techniques; D005804:Genes, Lethal; D006367:HeLa Cells; D006801:Humans; D017874:Immediate-Early Proteins / Q000037:antagonists & inhibitors / Q000235:genetics*; D030361:Papillomavirus Infections / Q000235:genetics* / Q000473:pathology; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000235:genetics*; D034741:RNA, Small Interfering / Q000378:metabolism*; D002583:Uterine Cervical Neoplasms / Q000235:genetics / Q000473:pathology / Q000821:virology*; D054462:p21-Activated Kinases / Q000037:antagonists & inhibitors / Q000235:genetics*; D000099184:Serum-Glucocorticoid Regulated Kinases
chemical_list: D017874:Immediate-Early Proteins; D017346:Protein Serine-Threonine Kinases; D034741:RNA, Small Interfering; D054462:p21-Activated Kinases; D000099184:Serum-Glucocorticoid Regulated Kinases; C515911:PAK3 protein, human