Abstract
Recent studies have shown that Beclin 1, a key regulator of autophagic process, is frequently downregulated and may serve as an independent prognostic biomarker for nonsmall cell lung cancer. However, the molecular mechanisms underlying its downregulation remain poorly understood. The signal transducer and activator of transcription 3 (Stat3) is a transcription factor which plays a crucial role for multiple tumor growth and progression. Here, we demonstrate that Beclin 1 is a direct transcriptional target of Stat3 in lung cancer cells. Interleukin-6 (IL-6) treatment or transfection of a constitutively activated Stat3 in AGS and NCI-H1650 cells inhibited Beclin 1 expression. At the molecular level, we further revealed that Stat3 could directly bind to the promoter region of Beclin 1 and repressed its transcription through recruiting histone deacetylase 3 (HDAC3). Collectively, our results suggest that the activated Stat3 may represent an important mechanism for Beclin 1 downregulation in nonsmall cell lung cancer development.
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Acknowledgments
We thank Dr. Yan Zuo-Qin for providing the CA-Stat3 plasmid.
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Supplementary Figure 1
Regulation of Beclin 1 by IL-6 in lung cancer cells. (A-B) Relative mRNA levels of Beclin 1 in AGS (A) and NCI-H1650 (B) cells treated with IL-6 (10 ng/ml) at different time as indicated. (GIF 2 kb)
Supplementary Figure 2
Regulation of Beclin 1 by dominant-negative Stat3 in lung cancer cells. (A-B) mRNA and protein levels of Beclin 1 were analyzed by Real-time PCR (A) and western blot (B) in AGS cells transfected with empty vector (EV) or a dominant-negative Stat3 mutant (DN-Stat3), for 24 or 36 hr, respectively. (C-D) mRNA and protein levels of Beclin 1 were analyzed by Real-time PCR (C) and western blot (D) in NCI-H1650 cells transfected with empty vector (EV) or a dominant-negative Stat3 mutant (DN-Stat3), for 24 or 36 hr, respectively. Plasmids containing green fluorescent protein (GFP) gene were co-transfected to evaluate the efficiency of transfection. (GIF 4 kb)
Supplementary Figure 3
Inhibitory roles of Stat3 on Beclin 1 expression relies on its recruitment of HDAC3. (A) mRNA levels of Beclin 1 in NCI-H1650 cells treated with vehicle control (PBS) or IL-6 (10ng/ml). Cells were pre-transfected with siRNA targeting HDAC3 or nonspecific negative controls (NC) for 24 hr. (B-C) mRNA levels of Beclin 1 in AGS cells treated with vehicle control (PBS) or IL-6 (10ng/ml). Cells were pre-transfected with siRNA targeting HDAC1 (B), HDAC2 (C) or nonspecific negative controls (NC) for 24 hr. Representative protein levels of HDAC1 and HDAC2 were displayed as indicated. (GIF 5 kb)
Supplementary Figure 4
Down-regulation of p53 by Stat3 in lung cancer cells. (A)Protein levels of p53 in AGS cells treated with vehicle control (PBS) or IL-6 (10ng/ml). Cells were pre-transfected with siRNA targeting HDAC3 or nonspecific negative controls (NC) for 24 hr. (B) Protein levels of p53 in AGS cells transfected with empty vector (EV) or constitutive activated Stat3 (CA-Stat3). Cells were pre-transfected with siRNA targeting HDAC3 or nonspecific negative controls (NC) for 24 hr. (GIF 3 kb)
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Miao, LJ., Huang, FX., Sun, ZT. et al. Stat3 inhibits Beclin 1 expression through recruitment of HDAC3 in nonsmall cell lung cancer cells. Tumor Biol. 35, 7097–7103 (2014). https://doi.org/10.1007/s13277-014-1961-6
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DOI: https://doi.org/10.1007/s13277-014-1961-6