Skip to main content
  • AACR Journals
    • 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
    • Meeting Abstracts
    • Collections
      • COVID-19 & Cancer Resource Center
      • Focus on Radiation Oncology
      • Novel Combinations
      • Reviews
      • 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 Journals
    • 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
Molecular Cancer Therapeutics
Molecular Cancer Therapeutics
  • Home
  • About
    • The Journal
    • AACR Journals
    • Subscriptions
    • Permissions and Reprints
  • Articles
    • OnlineFirst
    • Current Issue
    • Past Issues
    • Meeting Abstracts
    • Collections
      • COVID-19 & Cancer Resource Center
      • Focus on Radiation Oncology
      • Novel Combinations
      • Reviews
      • 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

Molecular Medicine in Practice

Exon 11 Skipping of E-Cadherin RNA Downregulates Its Expression in Head and Neck Cancer Cells

Sanjai Sharma, Wei Liao, Xiaofeng Zhou, David T.W. Wong and Alan Lichtenstein
Sanjai Sharma
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Wei Liao
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Xiaofeng Zhou
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
David T.W. Wong
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Alan Lichtenstein
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
DOI: 10.1158/1535-7163.MCT-11-0248 Published September 2011
  • Article
  • Figures & Data
  • Info & Metrics
  • PDF
Loading

Article Figures & Data

Figures

  • Additional Files
  • Figure 1.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 1.

    Aberrant transcripts in primary human tumor tissues. A, schematic showing the location of the probes to identify the wild-type and aberrant transcript (exon 11 skipped). B, different types of human cancer cDNA analyzed by real-time PCR analysis for wild-type and aberrant E-cadherin transcript (n = 9 for each tumor, n = 3 for matched organ-specific normal tissue). Black bars represent the percentage of tumor cDNAs with E-cadherin expression less than 50% of matched normal tissue; open bars represent the percentage of tumor cDNAs in which the percentage of aberrant transcript is at least 2-fold greater than the aberrant transcript in matched normal tissue. For the head and neck cancer group, the data show relative levels of total E-cadherin expression in 8 pairs of matched normal (N) and tumor tissues (T), relative to actin and adjusted to the matched normal control. C, head and neck cancer data from 8 matched groups is shown with wild type and percentage of exon 11–skipped transcript in each matched pair (log scale).

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

    Analysis of the aberrant transcript. A, RT-PCR analysis of untreated UMSCC12 cells treated with emetine or emetine plus actinomycin-D and analyzed for expression of E-cadherin RNA. With the same cDNA preparation FRS2 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were amplified. B and C, UMSCC14 and UMSCC12 cells were treated with actinomycin-D, and transcript-specific PCR conducted at different time points. D, relative expression of E-cadherin in tert keratinocytes, NHOK, and head and neck cell lines relative to expression in tert-kert cells. E, percentage exon 11–skipped transcript calculated by the cycle difference between the 2 transcripts with transcript-specific real-time PCR of untreated (E) and emetine treated, (F) cells.

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

    Minigene experiments show similar splicing patterns. A, schematic showing the minigene constructs with E-cadherin exon 10, 11, and 15. B, transient transfection and RT-PCR analysis (DNA gel) of UMSCC12 cells transfected with various minigene constructs. Primers bind to β−globin exon E1 and E3. Upper arrow indicates the correct expected size and the lower arrow indicates a transcript lacking exon 11. C, schematic for the location of probes for transcript-specific PCR. D, real-time PCR data showing the percentage aberrant transcript in tert-kert and head and neck cell lines (mean ± SD).

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

    Role of splicing factors in aberrant splicing. A, schematic with predicted binding sites for splicing factors around E-cadherin exon 11. B, real-time PCR data for expression of SFRS1, 2, 5, and 6 splicing factors in cell lines relative to tert-kert cells. C, expression of splicing factors by real-time PCR in primary human head and neck tumors relative to matched normal tissue (7 matched pairs).

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

    Effect of siRNA-mediated inactivation of SFRS2 on splicing. A, Western blot analysis for UMSCC12 and UMSCC22b cells transfected with SFRS2 siRNA or scrambled control sequence. B, bar diagram with percentage of aberrantly spliced transcript in cells transfected with exon 11 minigene construct and SFRS2 siRNA as determined by transcript-specific real-time PCR in UMSCC12 and UMSCC22b. Control cells were transfected with the exon 11 minigene construct and scrambled siRNA. Data are mean ± SD, n = 3. C, E-cadherin expression by real-time PCR analysis in 2 cell lines transfected with varying amounts of SFRS2 siRNA.

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

    Hypomethylating agent and aberrant splicing. A, methylation-specific PCR for UMSCC1 and UMSCC22b cell lines with and without azacytidine treatment. B, Western blot analysis for UMSCC1 and UMSCC22b showing E-cadherin expression and actin control with azacytidine treatment. C, expression of wild-type E-cadherin transcripts relative to the untreated cells and adjusted to actin, along with the percentage of aberrant transcript after azacytidine treatment.

Additional Files

  • Figures
  • Supplementary Data

    Files in this Data Supplement:

    • Supplementary Table 1 - XLS file - 13K, List of 100 top most downregulated genes in the array. Average of 5 cell H&N cell lines compared to Tert Kert cells
PreviousNext
Back to top
Molecular Cancer Therapeutics: 10 (9)
September 2011
Volume 10, Issue 9
  • 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 Molecular Cancer Therapeutics 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.
Exon 11 Skipping of E-Cadherin RNA Downregulates Its Expression in Head and Neck Cancer Cells
(Your Name) has forwarded a page to you from Molecular Cancer Therapeutics
(Your Name) thought you would be interested in this article in Molecular Cancer Therapeutics.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Citation Tools
Exon 11 Skipping of E-Cadherin RNA Downregulates Its Expression in Head and Neck Cancer Cells
Sanjai Sharma, Wei Liao, Xiaofeng Zhou, David T.W. Wong and Alan Lichtenstein
Mol Cancer Ther September 1 2011 (10) (9) 1751-1759; DOI: 10.1158/1535-7163.MCT-11-0248

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Share
Exon 11 Skipping of E-Cadherin RNA Downregulates Its Expression in Head and Neck Cancer Cells
Sanjai Sharma, Wei Liao, Xiaofeng Zhou, David T.W. Wong and Alan Lichtenstein
Mol Cancer Ther September 1 2011 (10) (9) 1751-1759; DOI: 10.1158/1535-7163.MCT-11-0248
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
    • Introduction
    • Materials and Methods
    • Results
    • Discussion
    • Disclosure of Potential Conflicts of Interest
    • Grant Support
    • Acknowledgments
    • Footnotes
    • References
  • Figures & Data
  • Info & Metrics
  • PDF
Advertisement

Related Articles

Cited By...

More in this TOC Section

  • EMT and CSCs in HER2+ Breast Cancer
  • Sorafenib Induces Reactive Oxygen Species Production
  • Methylglyoxal Enhances TRAIL Efficacy
Show more Molecular Medicine in Practice
  • Home
  • Alerts
  • Feedback
  • Privacy Policy
Facebook  Twitter  LinkedIn  YouTube  RSS

Articles

  • Online First
  • Current Issue
  • Past Issues
  • Meeting Abstracts

Info for

  • Authors
  • Subscribers
  • Advertisers
  • Librarians

About MCT

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

Copyright © 2021 by the American Association for Cancer Research.

Molecular Cancer Therapeutics
eISSN: 1538-8514
ISSN: 1535-7163

Advertisement