Steroid hormone control of cell death and cell survival: molecular insights using RNAi

Autor: J. Douglas Freeman, Melissa K. McConechy, Sharon M. Gorski, Lindsay DeVorkin, Ying-Chen Claire Hou, Suganthi Chittaranjan
Jazyk: angličtina
Rok vydání: 2009
Předmět:
Cancer Research
Programmed cell death
Ecdysone
lcsh:QH426-470
Cell Survival
medicine.medical_treatment
Biology
Cell morphology
Salivary Glands
Cell Line
03 medical and health sciences
chemistry.chemical_compound
0302 clinical medicine
RNA interference
Genetics
medicine
Developmental Biology/Developmental Molecular Mechanisms
Animals
Drosophila Proteins
Molecular Biology
Genetics (clinical)
Ecology
Evolution
Behavior and Systematics
030304 developmental biology
0303 health sciences
Cell Death
Cell growth
Genetics and Genomics/Functional Genomics
Cell Biology/Cellular Death and Stress Responses
Molecular biology
Cell biology
Steroid hormone
lcsh:Genetics
chemistry
Cell culture
Apoptosis
Drosophila
RNA Interference
Genetics and Genomics/Gene Discovery
030217 neurology & neurosurgery
Research Article
Zdroj: PLoS Genetics, Vol 5, Iss 2, p e1000379 (2009)
PLoS Genetics
ISSN: 1553-7404
1553-7390
Popis: The insect steroid hormone ecdysone triggers programmed cell death of obsolete larval tissues during metamorphosis and provides a model system for understanding steroid hormone control of cell death and cell survival. Previous genome-wide expression studies of Drosophila larval salivary glands resulted in the identification of many genes associated with ecdysone-induced cell death and cell survival, but functional verification was lacking. In this study, we test functionally 460 of these genes using RNA interference in ecdysone-treated Drosophila l(2)mbn cells. Cell viability, cell morphology, cell proliferation, and apoptosis assays confirmed the effects of known genes and additionally resulted in the identification of six new pro-death related genes, including sorting nexin-like gene SH3PX1 and Sox box protein Sox14, and 18 new pro-survival genes. Identified genes were further characterized to determine their ecdysone dependency and potential function in cell death regulation. We found that the pro-survival function of five genes (Ras85D, Cp1, CG13784, CG32016, and CG33087), was dependent on ecdysone signaling. The TUNEL assay revealed an additional two genes (Kap-α3 and Smr) with an ecdysone-dependent cell survival function that was associated with reduced cell death. In vitro, Sox14 RNAi reduced the percentage of TUNEL-positive l(2)mbn cells (p
Author Summary Hormones regulate complex signaling pathways required for the differentiation, growth, survival, and death of cells in diverse organisms. The insect steroid hormone 20-hydroxy ecdysone (ecdysone) triggers cell death of obsolete larval tissues, such as the midgut and salivary glands, during metamorphosis and provides a model system for understanding steroid hormone control of cell death and cell survival. Previous studies identified many genes and proteins associated with fruit fly salivary gland cell death, but functional verification was lacking. Here, we have analyzed 460 of those genes using RNAi, a genetic approach to inhibit gene function, to assess their possible cell death or cell survival related function. To our knowledge, this is the first large-scale functional screen for genes involved in steroid hormone regulated cell death and cell survival. We identified several novel ecdysone regulated components with a cell death/survival role, including genes with no previously known function. In vivo analyses of animals harboring an RNAi construct targeting the transcription factor Sox14, one of the genes identified, confirmed its role as a positive regulator of ecdysone-mediated cell death. Our results provide a foundation for further studies of the molecular mechanisms by which steroid hormones control the death and survival of cells.
Databáze: OpenAIRE
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