Abstract
Enhancer DNA methylation and expression of MYBPHL was studied in multiple myeloma (MM). By bisulfite genomic sequencing, among the three CpGs inside the MYBPHL enhancer, CpG1 was significantly hypomethylated in MM cell lines (6.7–50.0%) than normal plasma cells (37.5–75.0%) (P = 0.007), which was negatively correlated with qPCR-measured MYBPHL expression. In RPMI-8226 and WL-2 cells, bearing the highest CpG1 methylation, 5-azadC caused enhancer demethylation and expression of MYBPHL. In primary samples, higher CpG1 methylation was associated with lower MYBPHL expression. By luciferase assay, luciferase activity was enhanced by MYBPHL enhancer compared with empty vector control, but reduced by site-directed mutagenesis of each CpG. RNA-seq data of newly diagnosed MM patients showed that MYBPHL expression was associated with t(11;14). MOLP-8 cells carrying t(11;14) express the highest levels of MYBPHL, and its knockdown reduced cellular proliferation and increased cell death. Herein, as a proof-of-concept, our data demonstrated that the MYBPHL enhancer, particularly CpG1, was hypomethylated and associated with increased MYBPHL expression in MM, which was implicated in myelomagenesis.
Highlights
Enhancer DNA methylation and expression of MYBPHL was studied in multiple myeloma (MM)
Data demonstrated that higher MYBPHL enhancer methylation, CpG1, correlated with lower MYBPHL expression. (c) In RPMI-8226 and WL-2 cells, which had high level of MYBPHL enhancer methylation, treatment with 5-azadC led to MYBPHL enhancer demethylation, as evidenced by Bisulfite genomic sequencing (BGS), with concomitant re-expression of MYBPHL transcript, as detected by qPCR. (d) In primary myeloma samples, higher methylation of MYBPHL enhancer, CpG1, correlated with lower MYBPHL expression
By bisulfite genomic sequencing (BGS), DNA methylation of the MYBPHL enhancer was studied in bisulfite-converted DNA of CD138-sorted normal plasma cells from healthy bone marrow donors (n = 7), human MM cell lines (HMCLs; n = 7), and an enzymatically methylated positive control DNA
Summary
Enhancer DNA methylation and expression of MYBPHL was studied in multiple myeloma (MM). In RPMI-8226 and WL-2 cells, bearing the highest CpG1 methylation, 5-azadC caused enhancer demethylation and expression of MYBPHL. Higher CpG1 methylation was associated with lower MYBPHL expression. As a proof-of-concept, our data demonstrated that the MYBPHL enhancer, CpG1, was hypomethylated and associated with increased MYBPHL expression in MM, which was implicated in myelomagenesis. MM is defined by the presence of ≥ 10% clonal bone marrow plasma cells associated with features of end-organ damages including hypercalcaemia, renal failure, anaemia, or bone lesions that known as CRAB. Data demonstrated that higher MYBPHL enhancer methylation, CpG1, correlated with lower MYBPHL expression. (d) In primary myeloma samples, higher methylation of MYBPHL enhancer, CpG1, correlated with lower MYBPHL expression. Data of qPCR were mean delta Ct values from three independent experiments with triplicate in each
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
