Biochemical and Biophysical Research Communications
ATF5 increases cisplatin-induced apoptosis through up-regulation of Cyclin D3 transcription in HeLa cells
Section snippets
Experimental procedures
Materials. Restriction enzymes, bovine calf serum, DMEM, Trizol reagent, LipofectAMINE reagent, and the expression vectors pcDNA3.0, pcDNA3.1 were purchased from Invitrogen. PGL3-Basic and pRL-CMV were from Promega. Hoechst33258, dimethyl sulfoxide, cisplatin, and etoposide were purchased from Sigma Chemical. The anti-ATF5 Ab was purchased from Abcam. The anti-Cyclin D3 Ab, anti-Cyclin D2 Ab, and anti-Cyclin D1 Ab were purchased from Pharmingen. The anti-E2F1 Ab and anti-GAPDH Ab were purchased
Ectopic expression of ATF5 increases apoptosis induced by Cisplatin in HeLa cells
To elucidate the role of ATF5 in the DNA damage agent-induced apoptosis, ATF5 expression vector or empty vector was transfected into HeLa cells (Fig. 1A). Next, we investigated the effect of ATF5 on apoptosis after cisplatin treatment for 12 h. As shown in Fig. 1B, ATF5 sensitized HeLa cells to cisplatin-induced apoptosis as indicated by fragmented and condensed nuclei, indicating the pro-apoptotic role of ATF5 in HeLa cells. This conclusion was further supported in Figs. 1C and D. The
Discussion
ATF5 is a member of a large family of transcription factors originally identified more than a decade ago [18]. Since then, additional ATFs that medicate cell apoptosis have been reported [19], [20], [21], [22]. In this report, we have described several observations that implicated the role of ATF5 in cisplatin-induced apoptosis in HeLa cells. (a) ATF5 was up-regulated in the apoptosis process induced by cisplatin, which has been widely used in cervical treatment [23]. (b) The ectopic expression
Acknowledgments
This work was supported by National Natural Scientific Foundation of China (30470442 and 30330320) and CNHLPP (2004BA711A19) and a grant from the Development of Science and Technology of Shanghai (02DJ14002).
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Cited by (17)
Nucleophosmin (NPM1/B23) interacts with activating transcription factor 5 (ATF5) protein and promotes proteasome- and caspase-dependent ATF5 degradation in hepatocellular carcinoma cells
2012, Journal of Biological ChemistryCitation Excerpt :Re-expression of ATF5 in HCC inhibits cell proliferation and induces G2/M arrest (5). ATF5 has been shown to be involved in other cellular processes such as differentiation of neural progenitor cells (6–8), repression of cAMP-induced transcription in JEG3 choriocarcinoma cells and PC12 pheochromocytoma cells (6, 9), regulation of apoptosis (10–17), and response to various types of cellular stresses (18–20). ATF5 is known to subject to multilayered regulation that includes transcriptional regulation by EBF1 (21), translational regulation that is controlled by phosphorylated eIF2 (20, 23), and post-translational regulation that involves phosphorylation (2), acetylation (13), and Cdc34-dependent ubiquitin-mediated proteolysis (9, 24, 25).
BCL-2 is a downstream target of ATF5 that mediates the prosurvival function of ATF5 in a cell type-dependent manner
2011, Journal of Biological ChemistryRegulation of the human CHOP gene promoter by the stress response transcription factor ATF5 via the AARE1 site in human hepatoma HepG2 cells
2010, Life SciencesCitation Excerpt :This suggested that up-regulation of ATF5 protein might be responsible for the up-regulation of CHOP expression. Wei et al. (2008) reported that ATF5 protein was up-regulated by cisplatin, and its overexpression increased cisplatin-induced apoptosis in HeLa cells (Wei et al. 2006). On the other hand, it has been reported that in an interleukin 3-dependent cell line, ATF5 suppresses apoptosis resulting from cytokine deprivation (Persengiev et al. 2002), promotes cell survival through transcriptional activation of Hsp27 in a rat embryonal cardiac myoblast cell line, H9c2 (Wang et al. 2007), and suppresses the transactivational activity of p53 and blocks the p53-dependent apoptosis induced by ionizing irradiation in a fibrosarcoma cell line (Nishioka et al. 2009).
1,25(OH)<inf>2</inf>vitamin D<inf>3</inf> inhibits cell proliferation by promoting cell cycle arrest without inducing apoptosis and modifies cell morphology of mesenchymal multipotent cells
2010, Journal of Steroid Biochemistry and Molecular BiologyCdc34-mediated degradation of ATF5 is blocked by cisplatin
2008, Journal of Biological ChemistryCitation Excerpt :Immunofluorescence and confocal assay were performed as previously described (22). Reverse Transcription-PCR and Co-immunoprecipitations—Reverse transcription-PCR was performed as described previously (5). Primers used for PCR were as follows: ATF5 sense, 5′-AAGTCGGCGGCTCTGAGGTA-3′, and ATF5 antisense, 5′-GGACTCTGCCCGTTCCTTCA-3′; P21 sense, 5′-GACACCACTGGAGGGTGACT-3′, and P21 antisense, 5′-GGCGTTTGGAGTGGTAGAAA-3′.
ATF5 promotes cell survival through transcriptional activation of Hsp27 in H9c2 cells
2007, Cell Biology InternationalCitation Excerpt :Taken together, our results showed that ATF5 could promote H9c2 cell survival during heat shock stress. This is not consistent with the report (Wei et al., 2006) that ATF5 was a pro-apoptotic factor in cisplatin treated HeLa cells. The pro-apoptotic or anti-apoptotic ability of ATF5 in response to different extracellular stimulation indicated that ATF5 may act as a negative regulator or/and positive regulator depending on promoter context.