A Genetically Defined Normal Human Somatic Cell System to Study Ras Oncogenesis In Vivo and In Vitro

O’Hayer, Kevin; Counter, Christopher M.

doi:10.1016/s0076-6879(05)07050-3

Cited by 48 publications

(66 citation statements)

References 30 publications

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“…In humans, normal somatic cells can be driven to a tumorigenic state via the enforced expression of viral proteins that disrupt the tumor suppressors p53 and Rb and activate the proto-oncoprotein c-Myc, in conjunction with the mammalian oncogenic protein Ras and the hTERT telomerase catalytic subunit (Hahn et al, 1999;O'Hayer and Counter, 2006), all reflecting alterations commonly found in human cancers (Bos, 1989;Weinberg, 1991;Levine, 1997;Nesbit et al, 1999;Shay and Wright, 2002). More recently, we and others have demonstrated that expression of mammalian proteins can achieve the same feat (Kendall et al, 2006).…”

Section: Generation Of Porcine Cells Harboring Genetic Changes Commonmentioning

confidence: 88%

Genetic induction of tumorigenesis in Swine

et al. 2006

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The transition from basic to clinical cancer research for a number of experimental therapeutics is hampered by the lack of a genetically malleable, large animal model. To this end, we genetically engineered primary porcine cells to be tumorigenic by expression of proteins known to perturb pathways commonly corrupted in human cancer. Akin to human cells, these porcine cells were quite resistant to transformation, requiring multiple genetic changes. Moreover, the transformed porcine cells produced tumors when returned to the isogenic host animal. The ability to now rapidly and reproducibly genetically induce tumors of sizes similar to those treated clinically in a large mammal similar to humans in many respects will provide a robust cancer model for preclinical studies dependant on generating large tumors.

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Section: Generation Of Porcine Cells Harboring Genetic Changes Commonmentioning

confidence: 88%

Genetic induction of tumorigenesis in Swine

et al. 2006

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“…HEK-tTH cells were derived as described in Ref. 24. MEFs from cPLA2 Ϫ/Ϫ knock-out mice were obtained from J. Bonaventre (Harvard Medical School) and have been previously described (25).…”

Section: Methodsmentioning

confidence: 99%

“…All cells were maintained at 37°C in a 5% CO 2 incubator. Primary embryonic kidney cell lines were immortalized as previously described (24). C. trachomatis serovar LGV-L2 was propagated and stored as described in Ref.…”

Section: Methodsmentioning

confidence: 99%

“…For retroviral production, the pBabe retroviral system was used (27) and used as outlined in O'Hayer and Counter (24). Briefly, packaging plasmid (pCL-10A1) and transfer vector (pBabe) were transfected in a 1:1 ratio using Fugene6 (Roche) into 293T cells.…”

Section: Methodsmentioning

confidence: 99%

“…Human embryonic kidney (HEK) cells were immortalized by retroviral delivery of SV40 large and small T-antigen (T/t), and the catalytic domain of human telomerase (hTert), (24) and tested for the activation of RAS during infection. As with HeLa cells, infection of HEKt/TH cells led to increases in RAS-GTP and phosphorylation of ERK (Fig.…”

Section: Chlamydia Infection Leads To Broad Activation Of Ras Gtpases Inmentioning

confidence: 99%

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cPLA2 Regulates the Expression of Type I Interferons and Intracellular Immunity to Chlamydia trachomatis

Vignola

Kashatus

Taylor

et al. 2010

Journal of Biological Chemistry

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Infection with the obligate bacterial intracellular pathogenChlamydia trachomatis leads to the sustained activation of the small GTPase RAS and many of its downstream signaling components. In particular, the mitogen-activated protein kinase ERK and the calcium-dependent phospholipase cPLA 2 are activated and are important for the onset of inflammatory responses. In this study we tested if activation of ERK and cPLA 2 occurred as a result of RAS signaling during infection and determined the relative contribution of these signaling components to chlamydial replication and survival. We provide genetic and pharmacological evidence that during infection RAS, ERK, and, to a lesser extent, cPLA 2 activation are uncoupled, suggesting that Chlamydia activates individual components of this signaling pathway in a non-canonical manner. In human cell lines, inhibition of ERK or cPLA 2 signaling did not adversely impact C. trachomatis replication. In contrast, in murine cells, inhibition of ERK and cPLA 2 played a significant protective role against C. trachomatis. We determined that cPLA 2 -deficient murine cells are permissive for C. trachomatis replication because of their impaired expression of ␤ interferon and the induction of immunity-related GTPases (IRG) important for the containment of intracellular pathogens. Furthermore, the MAPK p38 was primarily responsible for cPLA 2 activation in Chlamydia-infected cells and IRG expression. Overall, these findings define a previously unrecognized role for cPLA 2 in the induction of cell autonomous cellular immunity to Chlamydia and highlight the many non-canonical signaling pathways engaged during infection.

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