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. 2017 Jun 20;7:3896. doi: 10.1038/s41598-017-04248-w

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Epigenetic regulation of tandem genes in lung cancer. (A) Outline of the tandem oriented reporter gene construct analyzed by chromatin immuno-precipitation (ChIP). The analyzed TO/EGFP and RASSF1A/RLuc promoter regions are depicted. For details see Fig. 5. (B) TREx293 cells stably transfected with TO-EGFP-2.3-RLuc (clone B7) were induced with 4 mM Dox (+) or uninduced (−) for four days (d) and histone modifications were analyzed by ChIP. Chromatin was precipitated with anti histone H3K9ac, histone H4ac and IgG antibodies (negative control) antibodies. DNA was isolated and analyzed by qPCR. Levels are plotted relative to 1% of input sample. (C) Expression of HDAC1, HDAC2, HDAC5, HDAC6, DNMT1, DNMT3A, DNMT3B and DNMT3L was analyzed by microarray in 49 primary squamous cell lung cancer (SCCa), 41 primary lung adenocarcinoma (AdCa), 15 non malignant lung disease samples (LD) and 6 normal lung tissues (NL)². Data are depicted as log2 transformed expression. (D) CPSF1 induced repression of RASSF1A. CPSF1 was cloned in pCMV-Tag1 and overexpressed in A427 cells. Endogenous RASSF1A level were analyzed by qRT-PCR and plotted relative to the empty control vector (=1). (E) Expression levels of CPSF1 in SCCa, AdCa, LD and NL. *p < 0.05, **p < 0.01 and ***p < 0.001.