title: Expulsion of micronuclei containing amplified genes contributes to a decrease in double minute chromosomes from malignant tumor cells.
abstract: Double minute chromosomes (DMs) are a hallmark of gene amplification. The relationship between the formation of DMs and the amplification of DM-carried genes remains to be clarified. The human colorectal cancer cell line NCI-H716 and human malignant primitive neuroectodermal tumor cell line SK-PN-DW are known to contain many DMs. To examine the amplification of DM-carried genes in tumor cells, we performed Affymetrix SNP Array 6.0 analyses and verified the regions of amplification in NCI-H716 and SK-PN-DW tumor cells. We identified the amplification regions and the DM-carried genes that were amplified and overexpressed in tumor cells. Using RNA interference, we downregulated seven DM-carried genes, (NDUFB9, MTSS1, NSMCE2, TRIB1, FAM84B, MYC and FGFR2) individually and then investigated the formation of DMs, the amplification of the DM-carried genes, DNA damage and the physiological function of these genes. We found that suppressing the expression of DM-carried genes led to a decrease in the number of DMs and reduced the amplification of the DM-carried genes through the micronuclei expulsion of DMs from the tumor cells. We further detected an increase in the number of γH2AX foci in the knockdown cells, which provides a strong link between DNA damage and the loss of DMs. In addition, the loss of DMs and the reduced amplification and expression of the DM-carried genes resulted in a decrease in cell proliferation and invasion ability.
authors.affiliation: Laboratory of Medical Genetics, Harbin Medical University, Harbin, People's Republic of China.
authors.forename: Wei
authors.initials: W
authors.lastname: Ji
authors.forename: Zehua
authors.initials: Z
authors.lastname: Bian
authors.forename: Yang
authors.initials: Y
authors.lastname: Yu
authors.forename: Chao
authors.initials: C
authors.lastname: Yuan
authors.forename: Yang
authors.initials: Y
authors.lastname: Liu
authors.forename: Lisa
authors.initials: L
authors.lastname: Yu
authors.forename: Chunxiang
authors.initials: C
authors.lastname: Li
authors.forename: Jing
authors.initials: J
authors.lastname: Zhu
authors.forename: Xueyuan
authors.initials: X
authors.lastname: Jia
authors.forename: Rongwei
authors.initials: R
authors.lastname: Guan
authors.forename: Chunyu
authors.initials: C
authors.lastname: Zhang
authors.forename: Xiangning
authors.initials: X
authors.lastname: Meng
authors.forename: Yan
authors.initials: Y
authors.lastname: Jin
authors.forename: Jing
authors.initials: J
authors.lastname: Bai
authors.forename: Jingcui
authors.initials: J
authors.lastname: Yu
authors.forename: Ki-Young
authors.initials: KY
authors.lastname: Lee
authors.forename: Wenjing
authors.initials: W
authors.lastname: Sun
authors.forename: Songbin
authors.initials: S
authors.lastname: Fu
mesh_terms: D014408:Biomarkers, Tumor / Q000235:genetics* / Q000378:metabolism; D015153:Blotting, Western; D002453:Cell Cycle; D002465:Cell Movement; D002467:Cell Nucleus / Q000235:genetics; D049109:Cell Proliferation; D002877:Chromosomes, Human / Q000235:genetics*; D015179:Colorectal Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D004249:DNA Damage / Q000235:genetics; D005455:Fluorescent Antibody Technique; D005784:Gene Amplification; D020869:Gene Expression Profiling; D006657:Histones / Q000235:genetics / Q000378:metabolism; D006801:Humans; D048629:Micronuclei, Chromosome-Defective; D018242:Neuroectodermal Tumors, Primitive / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D020411:Oligonucleotide Array Sequence Analysis; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured
chemical_list: D014408:Biomarkers, Tumor; C468783:H2AX protein, human; D006657:Histones; D012333:RNA, Messenger
keywords: double minute chromosomes; gene amplification; micronuclei; tumor
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