title: Met promotes the formation of double minute chromosomes induced by Sei-1 in NIH-3T3 murine fibroblasts.
abstract: BACKGROUND
  Sei-1 is an oncogene capable of inducing double minute chromosomes (DMs) formation. DMs are hallmarks of amplification and contribute to oncogenesis. However, the mechanism of Sei-1 inducing DMs formation remains unelucidated.
  
  
  RESULTS
  DMs formation significantly increased during serial passage in vivo and gradually decreased following culture in vitro. micro nuclei (MN) was found to be responsible for the reduction. Of the DMs-carrying genes, Met was found to be markedly amplified, overexpressed and highly correlated with DMs formation. Inhibition of Met signaling decreased the number of DMs and reduced the amplification of the DMs-carrying genes. We identified a 3.57Mb DMs representing the majority population, which consists of the 1.21 Mb AMP1 from locus 6qA2 and the 2.36 Mb AMP2 from locus 6qA2-3.
  
  
  METHODS
  We employed NIH-3T3 cell line with Sei-1 overexpression to monitor and characterize DMs in vivo and in vitro. Array comparative genome hybridization (aCGH) and fluorescence in situ hybridization (FISH) were performed to reveal amplification regions and DMs-carrying genes. Metaphase spread was prepared to count the DMs. Western blot and Met inhibition rescue experiments were performed to examine for involvement of altered Met signaling in Sei-1 induced DMs. Genomic walking and PCR were adopted to reveal DMs structure.
  
  
  CONCLUSIONS
  Met is an important promotor of DMs formation.
authors: Laboratory of Medical Genetics, Harbin Medical University, Harbin, China.
authors: Yantao
authors: Y
authors: Bao
authors: Laboratory of Medical Genetics, Harbin Medical University, Harbin, China.
authors: Jia
authors: J
authors: Liu
authors: Laboratory of Medical Genetics, Harbin Medical University, Harbin, China.
authors: Jia
authors: J
authors: You
authors: Laboratory of Medical Genetics, Harbin Medical University, Harbin, China.
authors: Di
authors: D
authors: Wu
authors: Laboratory of Medical Genetics, Harbin Medical University, Harbin, China.|Department of Genetics and Eugenics, Maternity and Child Care Center of Qinghuangdao, Qinghuangdao, China.
authors: Yang
authors: Y
authors: Yu
authors: Laboratory of Medical Genetics, Harbin Medical University, Harbin, China.
authors: Chang
authors: C
authors: Liu
authors: Laboratory of Medical Genetics, Harbin Medical University, Harbin, China.|Genetic Diagnosis Center, First People's Hospital of Yunnan Province, Yunnan, China.
authors: Lei
authors: L
authors: Wang
authors: Laboratory of Medical Genetics, Harbin Medical University, Harbin, China.
authors: Fei
authors: F
authors: Wang
authors: Laboratory of Medical Genetics, Harbin Medical University, Harbin, China.
authors: Lu
authors: L
authors: Xu
authors: Laboratory of Medical Genetics, Harbin Medical University, Harbin, China.
authors: Liqun
authors: L
authors: Wang
authors: Laboratory of Medical Genetics, Harbin Medical University, Harbin, China.
authors: Nan
authors: N
authors: Wang
authors: Laboratory of Medical Genetics, Harbin Medical University, Harbin, China.
authors: Xing
authors: X
authors: Tian
authors: Laboratory of Medical Genetics, Harbin Medical University, Harbin, China.
authors: Falin
authors: F
authors: Wang
authors: Laboratory of Medical Genetics, Harbin Medical University, Harbin, China.
authors: Hongbin
authors: H
authors: Liang
authors: Laboratory of Medical Genetics, Harbin Medical University, Harbin, China.
authors: Yating
authors: Y
authors: Gao
authors: Laboratory of Medical Genetics, Harbin Medical University, Harbin, China.
authors: Xiaobo
authors: X
authors: Cui
authors: Laboratory of Medical Genetics, Harbin Medical University, Harbin, China.
authors: Guohua
authors: G
authors: Ji
authors: Laboratory of Medical Genetics, Harbin Medical University, Harbin, China.
authors: Jing
authors: J
authors: Bai
authors: Scientific Research Centre, Second Affiliated Hospital, Harbin Medical University, Harbin, China.
authors: Jingcui
authors: J
authors: Yu
authors: Laboratory of Medical Genetics, Harbin Medical University, Harbin, China.
authors: Xiangning
authors: X
authors: Meng
authors: Laboratory of Medical Genetics, Harbin Medical University, Harbin, China.
authors: Yan
authors: Y
authors: Jin
authors: Laboratory of Medical Genetics, Harbin Medical University, Harbin, China.
authors: Wenjing
authors: W
authors: Sun
authors: Department of Clinical Oncology, Faculty of Medicine, The University of Hong Kong, Hong Kong, China.|State Key Laboratory of Oncology in South China and Collaborative Innovation Center for Cancer Medicine, Sun Yat-sen University Cancer Center, Guangzhou, China.
authors: Xin-Yuan
authors: XY
authors: Guan
authors: Laboratory of Medical Genetics, Harbin Medical University, Harbin, China.
authors: Chunyu
authors: C
authors: Zhang
authors: Laboratory of Medical Genetics, Harbin Medical University, Harbin, China.|Key Laboratory of Medical Genetics, Harbin Medical University, Heilongjiang Higher Education Institutions, Harbin, China.
authors: Songbin
authors: S
authors: Fu
mesh_terms: D000818:Animals; D002467:Cell Nucleus; D002471:Cell Transformation, Neoplastic; D002869:Chromosome Aberrations; D002875:Chromosomes / Q000648:ultrastructure*; D055028:Comparative Genomic Hybridization; D005784:Gene Amplification; D016678:Genome; D017404:In Situ Hybridization, Fluorescence; D008677:Metaphase; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D015162:Micronucleus Tests; D041681:NIH 3T3 Cells; D009687:Nuclear Proteins / Q000235:genetics*; D009857:Oncogenes; D051736:Proto-Oncogene Proteins c-mdm2 / Q000235:genetics; D019859:Proto-Oncogene Proteins c-met / Q000235:genetics*; D015398:Signal Transduction; D015534:Trans-Activators / Q000235:genetics*; D014157:Transcription Factors; D000092468:Bromodomain Containing Proteins
chemical_list: D009687:Nuclear Proteins; D051736:Proto-Oncogene Proteins c-mdm2; D019859:Proto-Oncogene Proteins c-met; D015534:Trans-Activators; D014157:Transcription Factors; C485730:Sertad1 protein, mouse; D000092468:Bromodomain Containing Proteins; C497207:Mdm2 protein, mouse
keywords: DMs; Sei-1; amplification; in vivo passage; met