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Related Experiment Videos

Gene expression studies in prostate cancer tissue: which reference gene should be selected for normalization?

Falk Ohl1, Monika Jung, Chuanliang Xu

  • 1Department of Urology, Charité-Universitätsmedizin Berlin, Campus Mitte, Schumannstrasse 20/21, 10098 Berlin, Germany. klaus.jung@charite.de

Journal of Molecular Medicine (Berlin, Germany)
|October 8, 2005
PubMed
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Selecting stable reference genes is crucial for accurate gene profiling in prostate cancer research. This study identified HPRT1, ALAS1, and K-ALPHA-1 as the most reliable genes for normalization in prostate cancer tissues.

Area of Science:

  • Molecular Biology
  • Oncology
  • Genetics

Background:

  • Quantitative reverse transcription-polymerase chain reaction (RT-PCR) relies on reference genes for accurate gene expression normalization.
  • The stability of commonly used housekeeping genes in prostate cancer tissues remains insufficiently validated.
  • Accurate normalization is essential for reliable gene profiling in cancer research.

Purpose of the Study:

  • To identify the most stable reference genes for normalization in prostate cancer tissues.
  • To evaluate the suitability of 16 candidate reference genes using real-time RT-PCR.
  • To demonstrate the impact of reference gene selection on target gene normalization.

Main Methods:

  • Examined mRNA expression of 16 candidate reference genes in matched malignant and non-malignant prostate cancer tissues from 17 patients.

Related Experiment Videos

  • Utilized quantitative real-time RT-PCR to measure gene expression levels (cycle threshold values).
  • Applied geNorm and NormFinder software to determine reference gene stability.

    • Main Results:

    • ACTB, RPL13A, and HMBS exhibited significant expression differences between malignant and non-malignant tissues.
    • HPRT1, ALAS1, and K-ALPHA-1 were identified as the most stable reference genes by both geNorm and NormFinder.
    • Normalization of the target gene RECK varied significantly depending on the reference gene used, highlighting the importance of stable gene selection.

    Conclusions:

    • HPRT1, ALAS1, and K-ALPHA-1 are recommended as reliable reference genes for gene expression studies in prostate cancer.
    • Using a single stable gene (HPRT1) or a combination of stable genes (HPRT1, ALAS1, K-ALPHA-1) improves normalization reliability.
    • Employing unstable reference genes can lead to erroneous conclusions in prostate cancer gene profiling.