Histone demethylase KDM3A induced the expression levels of PDK1 via H3K9me1 and H3K9me2 demethylation. (IMAGE)
Caption
(A) The analysis results of gene correlation analysis from the LUNG CANCER EXPLORER database showed that KDM3A was positively correlated with PDK1. (B) The Starbase database also confirmed that there was a positive correlation between KDM3A and PDK1. (C) The mRNA and protein expression levels of KDM3A were tested by quantitative reverse transcription PCR and Western blotting in PC-9 and PC-9/G cells. (D) The protein expression levels of KDM3A were tested by Western blotting in PC-9, PC-9/OR, HCC827, and HCC827/OR cell lines. (E) After silencing KDM3A in PC-9/G cells, the sensitivity of the cells to gefitinib increased. (F) The apoptosis rates were determined by flow cytometry, and KDM3A knockdown in PC-9/G cells induced higher apoptosis rates. (G) KDM3A knockdown in PC-9/G cells reduced the expression levels of PDK1, while the expression levels of H3K9me1 and H3K9me2 were increased. (H) The ENCODE database showed there were binding peaks of H3K9me1, H3K9me2, and KDM3A at the promoter region of PDK1. (I) Chromatin immunoprecipitation assay was conducted to determine the binding sites of KDM3A at the promoter region of the PDK1 gene. Cross-linked and sheared chromatin was immunoprecipitated with anti-KDM3A/histone H3 antibody or IgG, and quantitative reverse transcription PCR was used to detect the enrichment percentages. Data were statistically analyzed with Student’s t-test, and values were shown as mean ± standard deviation. ∗P < 0.05 and ∗∗P < 0.01.
Credit
Zhihao Zhou, Ruike Zhang, Zhaoyang Zhang, Liyuan Zhang, Wei Wang, Wenjing Liu, Chunyang Zhang, Gen Lin, Weimiao Yu, Bo Xu, Lin Wang, Bing-Hua Jiang
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