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Expression of CD44 3'-untranslated region regulates endogenous microRNA functions in tumorigenesis and angiogenesis

Zina Jeyapalan1, Zhaoqun Deng, Tatiana Shatseva

  • 1Sunnybrook Research Institute, Sunnybrook Health Sciences Centre, Toronto M4N 3M5, Canada.

Nucleic Acids Research
|December 15, 2010
PubMed

Insights

The CD44 3'-untranslated region (UTR) antagonizes microRNAs (miRNAs), increasing CD44 and CDC42 translation. This non-coding RNA region inhibits cancer cell proliferation and tumor growth, offering new therapeutic targets.

Area of Science:

  • Molecular Biology
  • Cancer Research
  • Gene Regulation

Background:

  • The 3 eal-untranslated region (UTR) of messenger RNAs (mRNAs) is crucial for post-transcriptional gene regulation.
  • MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression by binding to target mRNAs, typically in their 3 eal-UTRs.
  • Dysregulation of miRNA activity is implicated in various diseases, including cancer.

Purpose of the Study:

  • To investigate the role of the CD44 3 eal-UTR in regulating miRNA function and its impact on cancer progression.
  • To determine if the CD44 3 eal-UTR can antagonize cytoplasmic miRNAs and affect the translation of CD44 and its downstream targets.
  • To evaluate the functional consequences of CD44 3 eal-UTR activity on breast cancer cell behavior and tumor growth.

Main Methods:

  • Utilized computational algorithms to predict miRNA binding sites on CD44 and CDC42 3 eal-UTRs.
  • Performed luciferase assays to validate miRNA binding to predicted sites.
  • Conducted cell function assays (proliferation, colony formation, apoptosis, angiogenesis) in MT-1 human breast cancer cells.
  • Employed western blotting, immunohistochemical staining, and siRNA assays to confirm molecular interactions and functional effects.

Main Results:

  • The CD44 3 eal-UTR was shown to antagonize cytoplasmic miRNAs, leading to increased translation of CD44 and CDC42 mRNA.
  • Expression of the CD44 3 eal-UTR inhibited proliferation, colony formation, and tumor growth in vitro and in vivo.
  • The CD44 3 eal-UTR modulated endothelial cell activities, promoted angiogenesis, induced tumor cell apoptosis, and enhanced sensitivity to Docetaxel.
  • Computational predictions of miR-216a, miR-330, and miR-608 binding to CD44 and CDC42 3 eal-UTRs were experimentally confirmed.

Conclusions:

  • The non-coding CD44 3 eal-UTR acts as a competing endogenous RNA (ceRNA), sequestering miRNAs and thereby inactivating their function.
  • This mechanism leads to the de-repression of target mRNAs, including CD44 and CDC42, promoting cancer progression.
  • The CD44 3 eal-UTR represents a potential therapeutic target for modulating miRNA activity in cancer treatment.

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