Highly specific and sensitive detection of malignancy in urine samples from patients with urothelial cancer by CD44v8-10/CD44v10 competitive RT-PCR

Hideaki Miyake, Isamu Okamoto, Isao Hiara, Kazuo Gohji, Kazuki Yamanaka, Soichi Arakawa, Sadao Kamidono, Hideyuki Saya

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

CD44 is a widely expressed cell surface adhesion molecule in which various isoforms arise from alternative RNA splicing mechanism. Overexpression of specific CD44 splice variant, i.e., CD44v8-10, has been found in several human malignancies and is considered to be associated with tumor progression and metastasis. We have demonstrated a novel molecular approach, CD44v8-10/CD44v10 competitive reverse transcription-polymerase chain reaction (CC-RT-PCR) for the detection of cancer cells overexpressing CD44v8-10 by the measurement of the transcriptional level of CD44v8-10 relative to that of CD44v10 (v8-10/v10 ratio). In this study, we initially examined the expression of CD44 splice variants in human urothelial cancers and their adjacent normal urinary tissues by RT-PCR. Any CD44 variant isoforms were barely detectable in normal urinary tissues, whereas CD44v8-10 was predominantly expressed in 23 of the 30 (77%) urothelial cancer specimens. We then applied CC-RT-PCR to spontaneously voided urine samples from patients with 80 urothelial cancer and 50 various benign urologic diseases. The CC-RTPCR analysis revealed that all of the samples associated with benign diseases presented a predominant expression of the CD44v10 transcript (the v8-10/v10 ratios = 0.00-0.87), whereas 62 of the 80 samples associated with urothelial cancers mostly expressed the CD44v8-10 transcript (the v8-10/v10 ratios > 1.00). In addition, the positivity rate obtained by the CC-RT-PCR analysis was high regardless of the pathological grade of the urothelial cancers, although the sensitivity of the cytological examination declined with decreasing tumor grade. Our findings suggest strongly that CC- RT-PCR is a non-invasive useful tool for the diagnosis of urine samples from patients with urothetial cancer.

Original languageEnglish
Pages (from-to)560-564
Number of pages5
JournalInternational Journal of Cancer
Volume79
Issue number6
DOIs
Publication statusPublished - 1998
Externally publishedYes

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Urine
Polymerase Chain Reaction
Neoplasms
Reverse Transcription
Protein Isoforms
Urologic Diseases
Alternative Splicing
Cell Adhesion Molecules
Neoplasm Metastasis

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Highly specific and sensitive detection of malignancy in urine samples from patients with urothelial cancer by CD44v8-10/CD44v10 competitive RT-PCR. / Miyake, Hideaki; Okamoto, Isamu; Hiara, Isao; Gohji, Kazuo; Yamanaka, Kazuki; Arakawa, Soichi; Kamidono, Sadao; Saya, Hideyuki.

In: International Journal of Cancer, Vol. 79, No. 6, 1998, p. 560-564.

Research output: Contribution to journalArticle

Miyake, Hideaki ; Okamoto, Isamu ; Hiara, Isao ; Gohji, Kazuo ; Yamanaka, Kazuki ; Arakawa, Soichi ; Kamidono, Sadao ; Saya, Hideyuki. / Highly specific and sensitive detection of malignancy in urine samples from patients with urothelial cancer by CD44v8-10/CD44v10 competitive RT-PCR. In: International Journal of Cancer. 1998 ; Vol. 79, No. 6. pp. 560-564.
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abstract = "CD44 is a widely expressed cell surface adhesion molecule in which various isoforms arise from alternative RNA splicing mechanism. Overexpression of specific CD44 splice variant, i.e., CD44v8-10, has been found in several human malignancies and is considered to be associated with tumor progression and metastasis. We have demonstrated a novel molecular approach, CD44v8-10/CD44v10 competitive reverse transcription-polymerase chain reaction (CC-RT-PCR) for the detection of cancer cells overexpressing CD44v8-10 by the measurement of the transcriptional level of CD44v8-10 relative to that of CD44v10 (v8-10/v10 ratio). In this study, we initially examined the expression of CD44 splice variants in human urothelial cancers and their adjacent normal urinary tissues by RT-PCR. Any CD44 variant isoforms were barely detectable in normal urinary tissues, whereas CD44v8-10 was predominantly expressed in 23 of the 30 (77{\%}) urothelial cancer specimens. We then applied CC-RT-PCR to spontaneously voided urine samples from patients with 80 urothelial cancer and 50 various benign urologic diseases. The CC-RTPCR analysis revealed that all of the samples associated with benign diseases presented a predominant expression of the CD44v10 transcript (the v8-10/v10 ratios = 0.00-0.87), whereas 62 of the 80 samples associated with urothelial cancers mostly expressed the CD44v8-10 transcript (the v8-10/v10 ratios > 1.00). In addition, the positivity rate obtained by the CC-RT-PCR analysis was high regardless of the pathological grade of the urothelial cancers, although the sensitivity of the cytological examination declined with decreasing tumor grade. Our findings suggest strongly that CC- RT-PCR is a non-invasive useful tool for the diagnosis of urine samples from patients with urothetial cancer.",
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AU - Miyake, Hideaki

AU - Okamoto, Isamu

AU - Hiara, Isao

AU - Gohji, Kazuo

AU - Yamanaka, Kazuki

AU - Arakawa, Soichi

AU - Kamidono, Sadao

AU - Saya, Hideyuki

PY - 1998

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