Novel functional MAR elements of double minute chromosomes in human ovarian cells capable of enhancing gene expression.
Double minute chromosomes or double minutes (DMs) are cytogenetic hallmarks of extrachromosomal genomic amplification and play a critical role in tumorigenesis. Amplified copies of oncogenes in DMs have been associated with increased growth and survival of cancer cells but DNA sequences in DMs which are mostly non-coding remain to be characterized. Following sequencing and bioinformatics analyses, we have found 5 novel matrix attachment regions (MARs) in a 682 kb DM in the human ovarian cancer cell line, UACC-1598. By electrophoretic mobility shift assay (EMSA), we determined that all 5 MARs interact with the nuclear matrix in vitro. Furthermore, qPCR analysis revealed that these MARs associate with the nuclear matrix in vivo, indicating that they are functional. Transfection of MARs constructs into human embryonic kidney 293T cells showed significant enhancement of gene expression as measured by luciferase assay, suggesting that the identified MARS, particularly MARs 1 to 4, regulate their target genes in vivo and are potentially involved in DM-mediated oncogene activation.
Laboratory of Medical Genetics, Harbin Medical University, Harbin, China.
Yan
Y
Jin
Zheng
Z
Liu
Wei
W
Cao
Xinying
X
Ma
Yihui
Y
Fan
Yang
Y
Yu
Jing
J
Bai
Feng
F
Chen
Jesusa
J
Rosales
Ki-Young
KY
Lee
Songbin
S
Fu
D002460:Cell Line; D002875:Chromosomes / Q000648:ultrastructure*; D005260:Female; D018628:Gene Dosage; D005786:Gene Expression Regulation; D006801:Humans; D045170:Matrix Attachment Regions / Q000235:genetics* / Q000502:physiology; D015530:Nuclear Matrix; D010053:Ovary / Q000166:cytology / Q000648:ultrastructure*; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D015533:Transcriptional Activation
D011518:Proto-Oncogene Proteins