Amplification and Overexpression of the EMS 1 Oncogene, a Possible Prognostic Marker, in Human Hepatocellular Carcinoma Bao-Zhu Yuan, Xiaoling Zhou,

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Amplification and Overexpression of the EMS 1 Oncogene, a Possible Prognostic Marker, in Human Hepatocellular Carcinoma Bao-Zhu Yuan, Xiaoling Zhou, Drazen B. Zimonjic, Marian E. Durkin, Nicholas C. Popescu The Journal of Molecular Diagnostics Volume 5, Issue 1, Pages 48-53 (February 2003) DOI: 10.1016/S1525-1578(10)60451-5 Copyright © 2003 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 1 Isolation of genomic DNA fragments amplified in HCC by representational difference analysis. Comparative genomic hybridization profile for chromosomes 1 and 11 in hepatocellular carcinoma 94–25 (a), and detection of high-level DNA amplification on chromosomes 1 and 11 of the same tumor by reverse chromosome painting (b). Southern blot analysis of amplified fragments obtained by RDA (c). Four RDA clones (L1–1, L1–2, L3–2, and LA-7) were radiolabeled and hybridized to Southern blots of BglII-digested genomic DNA from the 94–25 liver tumor (94–25T) and from the surrounding normal tissue (94–25N). The blots were re-hybridized to a b-actin probe as a control. The sizes of the bands detected by the probes are indicated. The Journal of Molecular Diagnostics 2003 5, 48-53DOI: (10.1016/S1525-1578(10)60451-5) Copyright © 2003 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 2 Detection of amplification and overexpression of the EMSI gene in HCC. PCR analysis of EMSI amplification in DNA from primary HCC (a) and HCC cell lines (b), with tyrosine hydroxylase (TH) as an internal control. Northern blot analysis of EMSI mRNA levels in HCC cell lines and in normal liver (c). DNA from two primary HCC, cases 17 and 21, displayed a 3.2- and 2.9-fold amplification, respectively, compared with the normal counterpart (a). None of the HCC cell lines showed amplification (b). There is a very faint expression of EMS1 gene in normal liver cells but higher expression in HCC cell lines (3- to 7-fold increase as compared with normal liver cells). The blot was reprobed with GAPDH cDNA (c). The Journal of Molecular Diagnostics 2003 5, 48-53DOI: (10.1016/S1525-1578(10)60451-5) Copyright © 2003 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 3 Fluorescence in situ hybridization of chromosomes derived from PCL/PRF/5 and SK-Hep-1 cell lines. The chromosomes were hybridized to an EMS1 genomic probe (a and c) and rehybridized to a chromosome 11 painting probe (b and d). The number of fluorescent signals obtained with the EMS1-specific probe corresponds to the number of copies of chromosome 11q. The Journal of Molecular Diagnostics 2003 5, 48-53DOI: (10.1016/S1525-1578(10)60451-5) Copyright © 2003 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions