Epigenetic Role of Histone Lysine Methyltransferase and Demethylase on the Expression of Transcription Factors Associated with the Epithelial-to-Mesenchymal Transition of Lung Adenocarcinoma Metastasis to the Brain

SIMPLE SUMMARY: The epithelial-to-mesenchymal transition (EMT) is an essential step for cancer metastasis to the brain. The transcription factors (TFs), which are associated with EMT, plays a major role during EMT. The objectives of this study were to investigate the epigenetic modification of EMT by regulating the expression of EMT-TFs during the metastasis of lung cancer into the brain. Several EMT-TFs such as Slug, Twist, ZEB1, and FOXC2 had higher immunoreactivity in brain metastasis than lung cancer. MLL4 (H3K4 methyltransferase) induces the expression of Slug, UTX (H3K36me3 demethylase) induces the expression of ZEB1, and EZH2 (H3K27 methyltransferase) suppressed the expression of Twist in the analysis of immunohistochemical staining and qRT-PCR for the 46 paired samples of lung cancer and its brain metastasis in individual patients. These results were also statistically significantly associated with the survival of the patients. ABSTRACT: Purpose: The objective of this study was to investigate the epigenetic role of histone lysine methylation/demethylation on the expression of epithelial-to-mesenchymal transition (EMT) associated transcriptional factors (TFs) during the metastasis of lung adenocarcinoma to the brain. Methods: Paired samples of lung adenocarcinoma and brain metastasis (BM) were analyzed in 46 individual patients. Both samples were obtained by surgical resection or biopsy of the lung and brain. The paraffin-fixed formalin-embedded samples were obtained from the pathology archives in our institute. In samples of lung adenocarcinoma and BM, immunohistochemical staining was performed for epithelial markers, mesenchymal markers, EMT-TFs, histone lysine methyltransferase and demethylase. Results: The immunoreactivity of EMT-TFs such as Slug (15.6% vs. 42.6%, p = 0.005), Twist (23.6% vs. 45.9%, p = 0.010) and ZEB1 (15.0% vs. 55.9%, p = 0.002) was increased in BM compared with that in lung adenocarcinoma. Epigenetic inducers such as H3K4 methyltransferase (MLL4, p = 0.018) and H3K36me3 demethylase (UTX, p = 0.003) were statistically increased, and epigenetic repressors such as EZH2 (H3K27 methyltransferase, p = 0.046) were significantly decreased in BM compared with those in lung adenocarcinoma. The expression of UTX-ZEB1 (R(2) linear = 1.204) and MLL4-Slug (R(2) linear = 0.987) was increased in direct proportion, and EZH2-Twist (R(2) linear = −2.723) decreased in reverse proportion. Conclusions: The results suggest that certain histone lysine methyltransferase/demethylase, such as MLL4, UTX, and EZH2, regulate the expression of EMT-TFs such as Slug, ZEB1, and Twist epigenetically, which may thereby influence cancer metastasis from the lung to the brain.