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Signaling and Regulation

Phosphatidylinositol-3-OH Kinase or RAS Pathway Mutations in Human Breast Cancer Cell Lines

Antoinette Hollestelle, Fons Elstrodt, Jord H.A. Nagel, Wouter W. Kallemeijn and Mieke Schutte
Antoinette Hollestelle
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Fons Elstrodt
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Jord H.A. Nagel
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Wouter W. Kallemeijn
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Mieke Schutte
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DOI: 10.1158/1541-7786.MCR-06-0263 Published February 2007
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  • FIGURE 1.
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    FIGURE 1.

    Identification of the PTEN 802insTAGG/834delCTTC mutation in cell line CAMA-1 by PCR amplification and sequencing of genomic DNA (bottom electropherogram). The wild-type PTEN gene sequence is shown for comparison (top electropherogram).

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    FIGURE 2.

    Analysis of PTEN transcriptional silencing through promoter methylation by cell culture in the presence (+) or absence (−) of 5-azacytidine. Reverse transcription-PCR amplification products are shown from seven mutant and three wild-type PTEN breast cancer cell lines, using primers specific for PTEN (top fragments) and the HPRT housekeeper (bottom fragments). These cell lines included the three cell lines without detectable PTEN expression, but there was no indication of PTEN promoter methylation.

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  • Table 1.

    Mutations Identified in the PTEN, PIK3CA, RAS, and BRAF Genes in Human Breast Cancer Cell Lines

    Breast Cancer Cell LineAffected GeneGene SequenceTranscript SequencePredicted Protein Effect*Oncogenic
    HCC1937†PTENHD Ex. 1-9‡Not detectableNo expressionYes
    MDA-MB-468†PTENIVS4+1G>T‡c.del210_253 (Ex. 4)A72fsX5Yes
    BT549†PTEN821delG‡821delGV275XYes
    EVSA-TPTEN951delACTT‡951delACTTT319XYes
    CAMA-1PTEN274G>C§274G>CD92HYes
    802insTAGG/Not detectableNo expressionYes
    834delCTTC§
    ZR-75-1†PTEN323T>G‡323T>GL108RLikely
    MDA-MB-415†PTEN407G>A‡407G>AC136YYes
    MDA-MB-453PTEN919G>A919G>AE307KLikely
    BT474†PIK3CA333G>CNAK111NYes
    BT20†PIK3CA1616C>GNAP539RYes
    BT483†PIK3CA1624G>ANAE542KYes
    MCF-7†PIK3CA1633G>ANAE545KYes
    MDA-MB-361†PIK3CA1633G>ANAE545KYes
    MDA-MB-361PIK3CA1700A>GNAK567RLikely
    BT20†PIK3CA3140A>GNAH1047RYes
    MDA-MB-453†PIK3CA3140A>GNAH1047RYes
    OCUB-FPIK3CA3140A>G‡NAH1047RYes
    SK-BR-5PIK3CA3140A>GNAH1047RYes
    SUM102PT†PIK3CA3140A>GNAH1047RYes
    SUM185PE†PIK3CA3140A>G‡NAH1047RYes
    SUM190PT†PIK3CA3140A>GNAH1047RYes
    T47D†PIK3CA3140A>GNAH1047RYes
    UACC893†PIK3CA3140A>GNAH1047RYes
    SUM159PT†PIK3CA3140A>TNAH1047LYes
    MDA-MB-134VI†KRAS34G>CNAG12RYes
    SK-BR-7KRAS34G>TNAG12CYes
    SUM229PEKRAS35G>ANAG12DYes
    MPE600KRAS35G>TNAG12VYes
    MDA-MB-231†KRAS38G>ANAG13DYes
    Hs578T†HRAS35G>ANAG12DYes
    SUM159PTHRAS35G>ANAG12DYes
    SK-BR-7NRAS182A>GNAQ61RYes
    ZR-75-30†BRAF977T>CNAI326TUnknown
    MDA-MB-231BRAF1391G>TNAG464VYes
    DU4475BRAF1799T>ANAV600EYes
    MDA-MB-435sBRAF1799T>ANAV600EYes
    • Abbreviations: HD, homozygous deletion; Ex., exon; IVS, intervening sequence; del, deletion; ins, insertion; NA, not analyzed.

    • ↵* Frameshift mutations are indicated by the first changed codon and the number of newly encoded codons, including premature termination codon X.

    • ↵† Cell lines were reported to be mutated in references (17, 26, 37, 45-49).

    • ↵‡ Mutations were homozygous based on sequence analysis and confirmed with polymorphic markers.

    • ↵§ Mutations are heterozygous but are located on different alleles.

  • Table 2.

    Mutational Activation of the PI3K and RAS Pathways Is Mutually Exclusive in Human Breast Cancer Cell Lines

    Breast Cancer Cell LinesPTENPIK3CAKRASBRAFHRASNRAS
    BT549V275X
    CAMA-1D92H
    EVSA-TT319X
    HCC1937no protein
    MDA-MB-415C136Y
    MDA-MB-468A72fsX5
    ZR-75-1L108R
    MDA-MB-453E307KH1047R
    BT20P539R/H1047R
    MDA-MB-361E545K/K567R
    BT474K111N
    BT483E542K
    MCF-7E545K
    OCUB-FH1047R
    SK-BR-5H1047R
    SUM102PTH1047R
    SUM185PEH1047R
    SUM190PTNAH1047R
    T47DH1047R
    UACC893H1047R
    SUM159PTH1047LG12D
    Hs578TG12D
    SK-BR-7G12CQ61R
    MDA-MB-134VIG12R
    MPE600G12V
    SUM229PEG12D
    MDA-MB-231G13DG464V
    MDA-MB-435sV600E
    DU4475V600E
    ZR-75-30I326T
    MDA-MB-157
    MDA-MB-175VII
    MDA-MB-330
    MDA-MB-436
    SK-BR-3
    SUM149PT
    SUM225CWNNANA
    SUM1315MO2
    SUM52PE
    UACC812
    Mutation rate8 of 38 (21%)14 of 39 (36%)5 of 40 (13%)4 of 40 (10%)2 of 40 (5%)1 of 40 (3%)
    • NOTE: Overview of mutations that were identified in 40 human breast cancer cell lines. The mutations are detailed in Table 1.

    • Abbreviation: NA, not analyzed.

Additional Files

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    Files in this Data Supplement:

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Molecular Cancer Research: 5 (2)
February 2007
Volume 5, Issue 2
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Phosphatidylinositol-3-OH Kinase or RAS Pathway Mutations in Human Breast Cancer Cell Lines
Antoinette Hollestelle, Fons Elstrodt, Jord H.A. Nagel, Wouter W. Kallemeijn and Mieke Schutte
Mol Cancer Res February 1 2007 (5) (2) 195-201; DOI: 10.1158/1541-7786.MCR-06-0263

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Phosphatidylinositol-3-OH Kinase or RAS Pathway Mutations in Human Breast Cancer Cell Lines
Antoinette Hollestelle, Fons Elstrodt, Jord H.A. Nagel, Wouter W. Kallemeijn and Mieke Schutte
Mol Cancer Res February 1 2007 (5) (2) 195-201; DOI: 10.1158/1541-7786.MCR-06-0263
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