Molecular Detection of Cancer Cells by Competitive Reverse Transcription-Polymerase Chain Reaction Analysis of Specific CD44 Variant RNAs

Okamoto, Isamu; Morisaki, Tsuyoshi; Sasaki, Ji Ichiro; Miyake, Hideaki; Matsumoto, Mitsuhiro; Suga, Moritaka; Ando, Masayuki; Saya, Hideyuki

doi:10.1093/jnci/90.4.307

Cited by 60 publications

(50 citation statements)

References 49 publications

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“…An abnormal CD44v8-v10 gene transcript was noted with high frequency in exfoliated cells in clinical diagnostic fluid specimens such as pleural effusion and urine. 5,22 Of 61 patients whose malignancy was missed on cytology (false negative), 11 had it diagnosed by RT-PCR for CD44v8-v10. 21 Overexpression of CD44v8-v10 containing mRNA has been used as a serum, urinary, and tissue marker for bladder cancer 21,22,38 and is an independent prognostic factor for colon cancer.…”

Section: Discussionmentioning

confidence: 99%

“…Its specificity was confirmed by Western blot using normal lymph node as a positive control. 5 Biotin block (Ventana) was applied 3 min. A secondary antibody (Dako LSAB Kit mouse/goat/ rabbit) was applied for 25 min.…”

Section: Immunohistochemistry For Cd44v10mentioning

confidence: 99%

“…Splice variants are expressed by some normal epithelial cells in a tissue-specific manner and CD44v10 is expressed by normal lymphocytes. 5 Cancers express novel variant isoforms, reflecting deregulated mRNA splicing. Alternative splicing to produce variant exons takes place in the membrane-proximal extracellular domain and the cytoskeletal tail, and the cell's microenvironment influences this process.…”

mentioning

confidence: 99%

“…15,16 Inclusion of single or contiguous variant exons has been described by RT-PCR and sequencing in many benign and cancer tissues. 5,6,[14][15][16][17][18][19][20][21][22][23] We, 11 and subsequently others, 12,13 showed that PC loses immunohistochemical expression of CD44s and some CD44v isoforms. CD44v6 was absent by Figure 1 Splice variant CD44 exons are situated in the extracellular domain.…”

mentioning

confidence: 99%

“…This form is detectable by competitive RT-PCR in bladder cancer tissue and cytologic specimens. 5,6,21,22 Clear cell renal cell carcinoma expresses this form in a minority of cases: 4/15 grade 1 and 2/15 grade 3. 19 Expression of this isoform has prognostic value in colon cancer.…”

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confidence: 99%

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Prostate cancer invasion is influenced more by expression of a CD44 isoform including variant 9 than by Muc18

Omara-Opyene

Qiu

Shah

et al. 2004

Laboratory Investigation

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The standard form of cell adhesion glycoprotein CD44 is a metastasis suppressor in prostate cancer. However, we previously showed by RT-PCR and Western blotting that cancer overexpresses unique CD44 variant v7-v10 isoforms. Muc18 is another cell adhesion marker reportedly overexpressed by prostate cancer. Matched frozen section-confirmed tumor and benign tissues were harvested from 10 prostatectomy specimens and tumor was microdissected from two lymph node metastases. Tissues were homogenized for RNA preparations, and RT-PCR was performed for the CD44v7-v10 sequence. In cultured prostate cancer cells, we caused RNA interference against CD44v9 and/or Muc18. We used PC3M cells and a derivative cell line called G s a, that constitutively expresses this G-protein and is more invasive. Lipofection was performed for a green fluorescent protein plasmid and for two 22-mer DNA fragments, cloned into a plasmid expression vector to generate hairpin, interfering dsRNA. Assays for invasion into Matrigel, a basement membrane matrix, were performed in 4-5 experiments. RT-PCR demonstrated expression of a 608 bp band representing CD44v7-v10 or a 638 bp band of Prostate cancer remains a public health problem of enormous magnitude; it is the second most common cause of cancer fatality among North American men and the most common in Great Britain. Only a minority of all cases invade locally and metastasize, thus requiring altered expression of cell adhesion molecules to allow tumor cell detachment, migration through a stromal matrix, and lymphovascular invasion.CD44, a family of transmembrane glycoproteins involved in homotypic cell, cell-matrix, and cellcytoskeletal interaction, holds promise as a prostate tumor marker. The extracellular domain of CD44 binds numerous matrix substituents: hyaluronic acid, heparin-affinity growth factors, vascular endothelial growth factor, p185 HER2 , epidermal growth factor, and hepatocyte growth factor. Its intracellular domain binds to ezrin, radixin, moesin and merlin, which interact with the cytoskeleton; notably, neutral endopeptidase 24.11 competes with CD44 for this interaction. 1 CD44 also binds directly to the cytoskeletal substituent ankyrin, thus determining cell and tissue architectural form. 2 CD44 is an oncodevelopmental protein, with roles in

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Section: Discussionmentioning

confidence: 99%

Section: Immunohistochemistry For Cd44v10mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Prostate cancer invasion is influenced more by expression of a CD44 isoform including variant 9 than by Muc18

Omara-Opyene

Qiu

Shah

et al. 2004

Laboratory Investigation

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

et al. 1998

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Detection of bladder carcinoma by combined testing of urine for hyaluronidase and cytokeratin 20 RNAs

Eissa

Kassim

Labib

et al. 2005

Cancer

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BACKGROUNDA new, sensitive, noninvasive method for the detection of urothelial carcinomas of the urinary bladder would open new possibilities in both the diagnosis and followup of patients.METHODSThis study included 228 patients diagnosed with bladder carcinoma, 68 patients with benign bladder lesions, and 44 healthy persons served as the control group. All were subjected to: serologic schistosomiasis antibody assay in serum, urine cytology, estimation of urine hyaluronic acid (HA) by enzyme‐linked immunosorbent assay, and detection of CK‐20 and hyaluronidase (HAase) by reverse transcription polymerase chain reaction (RT‐PCR) in urothelial cells from voided urine.RESULTSHA mean rank was higher in benign and malignant groups than in the healthy group (P < 0.0001) and was significantly related to tumor grade (P = 0.021). HA best‐cutoff, determined using receiver operating characteristic curve to discriminate between malignant and nonmalignant groups, was 58.5 units/mg protein at 85.8% sensitivity and 60.7% specificity. HAase RNA showed superior sensitivity (90.8%) over cytology (68.9%) and CK‐20 (78.1%) with specificity of 93.4%, 98.1% and 80.2%, respectively. The sensitivity reached 94.7% at a specificity of 91.5% when combined with CK‐20. All 4 of the investigated markers were related to grade at P <0.05. Whereas only HAase and CK‐20 were significantly related to stage (P < 0.05). As to schistosomiasis, only HAase RNA positivity was significantly associated (P = 0.038).CONCLUSIONSHAase RNA is a promising noninvasive test with high sensitivity and specificity in bladder carcinoma detection. Cancer 2005. © 2005 American Cancer Society.

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Molecular Detection of Cancer Cells by Competitive Reverse Transcription-Polymerase Chain Reaction Analysis of Specific CD44 Variant RNAs

Abstract: CC-RT-PCR analysis of CD44v8-10 expression could be an important adjunct to cytologic examination in cancer diagnosis, especially in detecting exfoliated cancer cells in pleural effusions and urine.

Cited by 60 publications

References 49 publications

Prostate cancer invasion is influenced more by expression of a CD44 isoform including variant 9 than by Muc18

Prostate cancer invasion is influenced more by expression of a CD44 isoform including variant 9 than by Muc18

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

Detection of bladder carcinoma by combined testing of urine for hyaluronidase and cytokeratin 20 RNAs

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