Biochemical and Biophysical Research Communications
G-protein γ subunit GNG11 strongly regulates cellular senescence
Section snippets
Materials and methods
Cell culture. Human embryonic lung fibroblast (TIG-7) and HeLa cells were obtained from Japanese Cancer Research Resources Cell Bank. The cells were cultured at 37 °C in plastic Petri dishes containing Dulbecco’s modified Eagle’s medium supplemented with 10% fetal calf serum under 5% CO2 and 95% [11]. TIG-7 cells enter senescence at 75 PDLs under the above growth conditions. Quiescent cells were obtained by culturing cells in medium containing 0.1% serum for 5 days.
Transfection assay. A
Induction of GNG-11 in fibroblast and HeLa cells
We made an almost complete catalogue of genes up-regulated upon addition of 5′-bromodeoxyuridine (BrdU) in HeLa cells using PCR-based suppression subtractive hybridization [9] and DNA microarray analysis [10]. BrdU is shown to induce senescence-associated genes similar to but far more clearly than other systems of senescence [9], [10], [11]. This catalogue contains genes involved in remodeling of extracellular matrix, signal transduction, cell-cycle regulation, proliferation, and
Discussion
In the present study, GNG11 was shown to work as a novel type of regulator in cellular senescence. Induction of GNG11 in both senescent cells and BrdU-induced senescent-like phenomena reflects its involvement in senescence pathway. GNG11 is also induced in response to other senescence-inducing agents such as excess thymidine (data not shown), further establishing its candidature as a senescence marker. However, the most novel feature of GNG11 is that it quickly responds to oxidative stress.
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