Sp17 gene expression in myeloma cells is regulated by promoter methylation

British Journal of Cancer
Zhiqing WangS H Lim

Abstract

The mechanisms underlying sperm protein 17 (Sp17) gene expression in myeloma cells remained unclear. Using reverse transcription-polymerase chain reaction (RT-PCR), Sp17 transcripts were detected in ARK-B, ARP-1, RPMI-8226 and KMS-11 but not in H929, IM-9, MM1-R and U266 cells. Using a panel of primer pairs in methylation-sensitive PCR to amplify overlapping gene segments, our screening studies showed that the HpaII sites at -359 and -350 are involved in the regulation of Sp17 gene expression. To confirm the differences in methylation status between Sp17-positive and Sp17-negative cell lines, KMS-11 cells (Sp17-positive) and IM-9 cells (Sp17-negative) were subjected to the more accurate method of bisulphite conversion. KMS-11 cells were more hypomethylated at these HpaII sites of exon 1 compared to IM-9 cells, indicating the association of hypomethylated promoter with Sp17 gene expression. In addition, the level of methylation at other CpG sites within the promoter sequence was also higher in IM-9 than KMS-11. Exon 1 was cloned into a reporter vector, pCAT*3 Enhancer. Chloramphenicol acetyl transferase (CAT) activity was restored in cells transfected with the recombinant plasmid, indicating the promoter function of exon 1. Expo...Continue Reading

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Citations

Oct 5, 2010·Wiley Interdisciplinary Reviews. Systems Biology and Medicine·Amy SharmaAmit Verma
Oct 13, 2006·International Journal of Cancer. Journal International Du Cancer·Zhiqing WangSeah H Lim
Jan 17, 2007·British Journal of Haematology·Farouk MeklatSeah H Lim
Nov 28, 2008·British Journal of Haematology·Farouk MeklatSeah H Lim
Sep 3, 2005·American Journal of Hematology·Alisher R DadabayevSeah H Lim
Sep 28, 2005·International Journal of Cancer. Journal International Du Cancer·Zhiqing WangSeah H Lim
Jul 1, 2006·Human Molecular Genetics·Michael KoslowskiOzlem Türeci
Oct 23, 2012·International Reviews of Immunology·Maurizio Chiriva-InternatiRead Alalawi
Mar 28, 2017·Cancer Immunology, Immunotherapy : CII·Rory M ShallisSeah H Lim
Nov 10, 2012·Tissue Antigens·M F Gjerstorff, H J Ditzel
May 13, 2008·Leukemia Research·Yana ZhangSeah H Lim
Oct 28, 2008·Methods : a Companion to Methods in Enzymology·Kyle R PomraningMichael Freitag

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Methods Mentioned

BETA
PCR
restriction digest
transfection
transfect
Assay
restriction

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