Met promotes the formation of double minute chromosomes induced by Sei-1 in NIH-3T3 murine fibroblasts
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Yantao Bao1,*, Jia Liu1,*, Jia You1, Di Wu1, Yang Yu1,2, Chang Liu1, Lei Wang1,3, Fei Wang1, Lu Xu1, Liqun Wang1, Nan Wang1, Xing Tian1, Falin Wang1, Hongbin Liang1, Yating Gao1, Xiaobo Cui1, Guohua Ji1, Jing Bai1, Jingcui Yu4, Xiangning Meng1, Yan Jin1, Wenjing Sun1, Xin-yuan Guan5,6, Chunyu Zhang1, Songbin Fu1,7
1Laboratory of Medical Genetics, Harbin Medical University, Harbin, China
2Department of Genetics and Eugenics, Maternity and Child Care Center of Qinghuangdao, Qinghuangdao, China
3Genetic Diagnosis Center, First People's Hospital of Yunnan Province, Yunnan, China
4Scientific Research Centre, Second Affiliated Hospital, Harbin Medical University, Harbin, China
5Department of Clinical Oncology, Faculty of Medicine, The University of Hong Kong, Hong Kong, China
6State Key Laboratory of Oncology in South China and Collaborative Innovation Center for Cancer Medicine, Sun Yat-sen University Cancer Center, Guangzhou, China
7Key Laboratory of Medical Genetics, Harbin Medical University, Heilongjiang Higher Education Institutions, Harbin, China
*These authors contributed equally to this work
Songbin Fu, email: firstname.lastname@example.org
Chunyu Zhang, email: email@example.com
Keywords: Sei-1, in vivo passage, DMs, amplification, met
Received: March 30, 2016 Accepted: July 19, 2016 Published: August 01, 2016
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.
Materials and 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.
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