Skip to main content
  • AACR Publications
    • Blood Cancer Discovery
    • Cancer Discovery
    • Cancer Epidemiology, Biomarkers & Prevention
    • Cancer Immunology Research
    • Cancer Prevention Research
    • Cancer Research
    • Clinical Cancer Research
    • Molecular Cancer Research
    • Molecular Cancer Therapeutics

AACR logo

  • Register
  • Log in
  • My Cart
Advertisement

Main menu

  • Home
  • About
    • The Journal
    • AACR Journals
    • Subscriptions
    • Permissions and Reprints
  • Articles
    • OnlineFirst
    • Current Issue
    • Past Issues
    • CCR Focus Archive
    • Meeting Abstracts
    • Collections
      • COVID-19 & Cancer Resource Center
      • Breast Cancer
      • Clinical Trials
      • Immunotherapy: Facts and Hopes
      • Editors' Picks
      • "Best of" Collection
  • For Authors
    • Information for Authors
    • Author Services
    • Best of: Author Profiles
    • Submit
  • Alerts
    • Table of Contents
    • Editors' Picks
    • OnlineFirst
    • Citation
    • Author/Keyword
    • RSS Feeds
    • My Alert Summary & Preferences
  • News
    • Cancer Discovery News
  • COVID-19
  • Webinars
  • Search More

    Advanced Search

  • AACR Publications
    • Blood Cancer Discovery
    • Cancer Discovery
    • Cancer Epidemiology, Biomarkers & Prevention
    • Cancer Immunology Research
    • Cancer Prevention Research
    • Cancer Research
    • Clinical Cancer Research
    • Molecular Cancer Research
    • Molecular Cancer Therapeutics

User menu

  • Register
  • Log in
  • My Cart

Search

  • Advanced search
Clinical Cancer Research
Clinical Cancer Research
  • Home
  • About
    • The Journal
    • AACR Journals
    • Subscriptions
    • Permissions and Reprints
  • Articles
    • OnlineFirst
    • Current Issue
    • Past Issues
    • CCR Focus Archive
    • Meeting Abstracts
    • Collections
      • COVID-19 & Cancer Resource Center
      • Breast Cancer
      • Clinical Trials
      • Immunotherapy: Facts and Hopes
      • Editors' Picks
      • "Best of" Collection
  • For Authors
    • Information for Authors
    • Author Services
    • Best of: Author Profiles
    • Submit
  • Alerts
    • Table of Contents
    • Editors' Picks
    • OnlineFirst
    • Citation
    • Author/Keyword
    • RSS Feeds
    • My Alert Summary & Preferences
  • News
    • Cancer Discovery News
  • COVID-19
  • Webinars
  • Search More

    Advanced Search

Cancer Therapy: Preclinical

Lapatinib, a Dual EGFR and HER2 Kinase Inhibitor, Selectively Inhibits HER2-Amplified Human Gastric Cancer Cells and is Synergistic with Trastuzumab In vitro and In vivo

Zev A. Wainberg, Adrian Anghel, Amrita J. Desai, Raul Ayala, Tong Luo, Brent Safran, Marlena S. Fejzo, J. Randolph Hecht, Dennis J. Slamon and Richard S. Finn
Zev A. Wainberg
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Adrian Anghel
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Amrita J. Desai
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Raul Ayala
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Tong Luo
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Brent Safran
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Marlena S. Fejzo
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
J. Randolph Hecht
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Dennis J. Slamon
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Richard S. Finn
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
DOI: 10.1158/1078-0432.CCR-09-1112 Published March 2010
  • Article
  • Figures & Data
  • Info & Metrics
  • PDF
Loading

Article Figures & Data

Figures

  • Tables
  • Fig. 1.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 1.

    A, growth-inhibitory effects of lapatinib in gastric and esophageal cancer cell lines. Cells were treated with lapatinib at ranges from 0.001 to 10 μmol/L. N87 and OE19 (two HER2-amplified cell lines) were the most sensitive cell lines to lapatinib. This data is the result of experiments repeated in triplicate. B, HER2 gene copy number was analyzed using FISH. Samples were counterstained with DAPI and scored as positive for amplification if they had either (A) greater than four copies of HER2/neu per chromosome 17 centromere in at least 20 cells, or had (B) uncountable clusters of signals suggestive of an amplicon in 20 or more cells. HER2/Neu amplification was seen in two cell lines: OE19 and N87. SNU1 is representative of nonamplification.

  • Fig. 2.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 2.

    Cell signaling after treatment with lapatinib and erlotinib in gastric cancer cell lines: all cell lines were treated with 1 μmol/L of lapatinib. A, no appreciable differences in EGFR expression were seen after treatment with lapatinib but pEGFR was decreased in the two most sensitive cell lines. No differences in total HER2 expression were seen after treatment with lapatinib but pHER2 was also decreased in the two most sensitive cell lines. B, downregulation of pAKT was seen in the most lapatinib-sensitive cell lines. Similarly, downregulation of pERK was seen in the most lapatinib-sensitive cell lines.

  • Fig. 3.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 3.

    Lapatinib induces G0-G1 cell cycle arrest and apoptosis in HER2-amplified gastric cancer. A, N87 and OE19 cells were treated with 1 μmol/L of lapatinib and cells were analyzed for DNA content by flow cytometry. The proportion of cells that undergo apoptosis is increased and is maintained both at 48 h and 5 d after treatment. B, N87 and OE19 both show an increase of the G0-G1 fraction after lapatinib treatment. No appreciable differences in the cell cycle are seen in SNU1 and SNU5 after treatment with lapatinib.

  • Fig. 4.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 4.

    The effects of lapatinib are distinct from the effects of anti-EGFR blockade by erlotinib or anti-HER2 blockade by trastuzumab. Cell signaling after treatment with erlotinib in gastric cancer cell lines: all cell lines were treated with 1 μmol/L of erlotinib. Some downregulation of pEGFR was seen in the most erlotinib-sensitive cell lines and no effect was seen on total EGFR expression. As in lapatinib-sensitive cells, downregulation of pERK was seen in the most erlotinib-sensitive cell lines. No effects in pHER2 or total HER2 expression were seen after treatment with erlotinib.

  • Fig. 5.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 5.

    The combination of lapatinib and trastuzumab is synergistic in HER2-amplified gastric cancer. A, in this experiment, the lapatinib concentrations ranged from 3.125 to 100 nmol/L for OE19 and 0.625 to 20 nmol/L for N87. For trastuzumab, doses ranged from 0.3125 to 10 μg/mL for both cell lines. Growth inhibition was calculated as a percentage of the untreated controls. Experiments were done in duplicate for each cell line. B, mean CI values for lapatinib-trastuzumab combinations in two HER2-amplified gastric cancer cell lines. Error bars indicate the 95% CI of the mean value. CI values were derived from variables of the median effects plots, and statistical tests were used to determine significance. In this analysis, synergy is defined as CI values significantly lower than 1.0, antagonism as CI values significantly higher than 1.0, and additivity as CI values equal to 1.0. Synergistic interactions were observed in both cell lines ranging from 0.228 (P < 0.0001) in OE19 to 0.226 (P < 0.0005) in N87. C, cells were treated with lapatinib at 100 nmol/L (OE19) or 20 nm (N87), trastuzumab (10 μg/mL) or the combination. Cells were analyzed for DNA content by flow cytometry. Both N87 and OE19 show a statistically significant increase in apoptosis with combination therapy than with either lapatinib or trastuzumab alone. There is also an increase in G0-G1 cell cycle arrest in the combination than with either lapatinib or trastuzumab alone. D, comparisons of signaling on downstream markers with lapatinib alone, trastuzumab alone or the combination. In this experiment, lapatinib treatment was at 100 nmol/L (OE19) or 20 nm (N87) combined with trastuzumab at 10 μg/mL. Minimal downregulation of pAKT, pERK are seen with lapatinib alone in both cell lines. Trastuzumab alone also has no effect on any of these markers. The combination of the two drugs causes a significant downregulation of pAKT and pERK in both HER2-amplified cell lines but is most pronounced in N87, the cell line with the highest level of amplification. No significant changes were seen with the combination in either pEGFR or pHER2 downregulation.

  • Fig. 6.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 6.

    Antitumor activity of lapatinib and trastuzumab on N87-bearing xenografts. N87 cells (5 × 106 cells with 50% Matrigel) were injected s.c. into nude mice and mice were randomized into one of four groups (n = 11/group). Treatment started at day 4 after injection with either lapatinib (100 mg/kg daily for 3 wk), trastuzumab (10 mg/kg i.p. twice weekly) or sterile PBS (i.p. twice weekly) or the combination of lapatinib and trastuzumab at the doses above. Tumors were measured biweekly. Student's t test was used to compare tumor sizes between the groups and results are presented as mean. Error bars represent SD of the mean. Student's t test was done at the end of the experiment. All comparisons were statistically significant between the following groups: combo versus L or T alone, P < 0.05; combo versus control, P < 0.001; L or T versus control, P < 0.001.

Tables

  • Figures
  • Table 1.

    Lapatinib and erlotinib concentrations that achieve IC50 and the corresponding k-Ras, and HER2 molecular status in gastric and esophageal cancer cells

    Cell lineLapatinib IC50 (mean ± SE, μmol/L)Erlotinib IC50 growth (mean ± SE, μmol/L)Trastuzumab, growth inhibition (%)HER2 amplification statusK-Ras mutation
    N870.01 ± 0.043.32 ± 0.3715.50 ± 6.08AmplifiedWT
    OE190.09 ± 0.022.31 ± 0.5034.53 ± 5.20AmplifiedWT
    NUGC40.35 ± 0.030.24 ± 0.040Not amplifiedWT
    NUGC32.24 ± 0.550.70 ± 0.010Not amplifiedWT
    FU974.86 ± 0.344.76 ± 0.960Not amplifiedWT
    SNU168.58 ± 0.696.50 ± 1.310Not amplifiedWT
    IM95>10>102.80Not amplifiedWT
    IM95m>10>107.85Not amplifiedWT
    MKN74>10>1010.60Not amplifiedWT
    MKN1>100.96 ± 0485.83Not amplifiedWT
    KATOIII>105.98 ± 0.984.18Not amplifiedWT
    AGS>10>105.01Not amplifiedG12D (exon 1)
    SNU1>10>100Not amplifiedG12D (exon 1)
    SNU5>10>100Not amplifiedWT

    NOTE: Fourteen gastric and esophageal cancer cell lines were treated with lapatinib, erlotinib, and trastuzumab as described. Lapatinib and erlotinib reported as concentrations that achieve IC50 whereas the effects of trastuzumab are reported as a percentage of growth inhibited. HER2 amplification status and k-Ras mutation status of the cell lines as measured by FISH and PCR, respectively.

PreviousNext
Back to top
Clinical Cancer Research: 16 (5)
March 2010
Volume 16, Issue 5
  • Table of Contents
  • Table of Contents (PDF)
  • About the Cover

Sign up for alerts

View this article with LENS

Open full page PDF
Article Alerts
Sign In to Email Alerts with your Email Address
Email Article

Thank you for sharing this Clinical Cancer Research article.

NOTE: We request your email address only to inform the recipient that it was you who recommended this article, and that it is not junk mail. We do not retain these email addresses.

Enter multiple addresses on separate lines or separate them with commas.
Lapatinib, a Dual EGFR and HER2 Kinase Inhibitor, Selectively Inhibits HER2-Amplified Human Gastric Cancer Cells and is Synergistic with Trastuzumab In vitro and In vivo
(Your Name) has forwarded a page to you from Clinical Cancer Research
(Your Name) thought you would be interested in this article in Clinical Cancer Research.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Citation Tools
Lapatinib, a Dual EGFR and HER2 Kinase Inhibitor, Selectively Inhibits HER2-Amplified Human Gastric Cancer Cells and is Synergistic with Trastuzumab In vitro and In vivo
Zev A. Wainberg, Adrian Anghel, Amrita J. Desai, Raul Ayala, Tong Luo, Brent Safran, Marlena S. Fejzo, J. Randolph Hecht, Dennis J. Slamon and Richard S. Finn
Clin Cancer Res March 1 2010 (16) (5) 1509-1519; DOI: 10.1158/1078-0432.CCR-09-1112

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Share
Lapatinib, a Dual EGFR and HER2 Kinase Inhibitor, Selectively Inhibits HER2-Amplified Human Gastric Cancer Cells and is Synergistic with Trastuzumab In vitro and In vivo
Zev A. Wainberg, Adrian Anghel, Amrita J. Desai, Raul Ayala, Tong Luo, Brent Safran, Marlena S. Fejzo, J. Randolph Hecht, Dennis J. Slamon and Richard S. Finn
Clin Cancer Res March 1 2010 (16) (5) 1509-1519; DOI: 10.1158/1078-0432.CCR-09-1112
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Jump to section

  • Article
    • Abstract
    • Materials and Methods
    • Results
    • Discussion
    • Disclosure of Potential Conflicts of Interest
    • Acknowledgments
    • Footnotes
    • References
  • Figures & Data
  • Info & Metrics
  • PDF
Advertisement

Related Articles

Cited By...

More in this TOC Section

  • Targeting HER2 with Osimertinib in NSCLC
  • Combined VEGF/EGFR Inhibition
  • Radiotherapy with IDO1/PD-1 Blockade Treats Advanced GBM
Show more Cancer Therapy: Preclinical
  • Home
  • Alerts
  • Feedback
  • Privacy Policy
Facebook  Twitter  LinkedIn  YouTube  RSS

Articles

  • Online First
  • Current Issue
  • Past Issues
  • CCR Focus Archive
  • Meeting Abstracts

Info for

  • Authors
  • Subscribers
  • Advertisers
  • Librarians

About Clinical Cancer Research

  • About the Journal
  • Editorial Board
  • Permissions
  • Submit a Manuscript
AACR logo

Copyright © 2021 by the American Association for Cancer Research.

Clinical Cancer Research
eISSN: 1557-3265
ISSN: 1078-0432

Advertisement