Abstract
Aim—To demonstrate the feasibility of studying specific gene expression in fine needle aspirates from clinical lesions. The reverse transcription/polymerase chain reaction (RT-PCR) technique was used to demonstrate CD44 gene expression in cells from diagnostic fine needle aspirates taken from patients attending the outpatient clinic for breast diseases.
Methods—Polyadenylated RNA was extracted from the cells remaining in the syringe barrel after fine needle aspirate cytological diagnosis of 41 patients with breast lesions. Analysis of CD44 gene expression was performed by RT-PCR using primers flanking the site for insertion of the variant exons. The resulting products were separated on 1·2% agarose gels, transferred to nylon membranes using Southern blotting and hybridised with specific probes for standard (constitutive) and variant exons of this gene.
Results—On hybridisation with the CD44 standard exon probe, the expected amplified product of approximately 482 base pairs was visualised in 22 of 41 samples examined. Further hybridisation with the “variant” exon probes (exons 7 (v2), 8 (v3), 9b (v4), 12 (v7), and 15 (v10)) on 12 of these samples showed the presence of large molecular variants in all of these samples. However, the expression pattern detected with the probes for exons 7 (v2), 8 (v3) and 9b (v4) differed among the patients.
Conclusions—Expression of the standard and variant regions of the CD44 gene in cells remaining in the syringe after fine needle aspiration was demonstrated using RT-PCR. The 5′ variant exon probes seemed to show different patterns of expression among the patients. Further studies are currently being conducted to determine whether there is any correlation between expression of the various components of this gene and cytological diagnosis. Using this method, it would be possible to study the expression of other candidate marker genes in breast cancer using fine needle aspirates.
Keywords: CD44
Keywords: fine needle aspirates
Keywords: breast cancer
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- Buley I. D. Update on special techniques in routine cytopathology. J Clin Pathol. 1993 Oct;46(10):881–885. doi: 10.1136/jcp.46.10.881. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Finn L., Dougherty G., Finley G., Meisler A., Becich M., Cooper D. L. Alternative splicing of CD44 pre-mRNA in human colorectal tumors. Biochem Biophys Res Commun. 1994 Apr 29;200(2):1015–1022. doi: 10.1006/bbrc.1994.1551. [DOI] [PubMed] [Google Scholar]
- Ihalainen J., Taavitsainen M., Salmivaara T., Palotie A. Diagnosis of pancreatic lesions using fine needle aspiration cytology: detection of K-ras point mutations using solid phase minisequencing. J Clin Pathol. 1994 Dec;47(12):1082–1084. doi: 10.1136/jcp.47.12.1082. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Joensuu H., Klemi P. J., Toikkanen S., Jalkanen S. Glycoprotein CD44 expression and its association with survival in breast cancer. Am J Pathol. 1993 Sep;143(3):867–874. [PMC free article] [PubMed] [Google Scholar]
- Kaufmann M., Heider K. H., Sinn H. P., von Minckwitz G., Ponta H., Herrlich P. CD44 variant exon epitopes in primary breast cancer and length of survival. Lancet. 1995 Mar 11;345(8950):615–619. doi: 10.1016/s0140-6736(95)90521-9. [DOI] [PubMed] [Google Scholar]
- Matsumura Y., Hanbury D., Smith J., Tarin D. Non-invasive detection of malignancy by identification of unusual CD44 gene activity in exfoliated cancer cells. BMJ. 1994 Mar 5;308(6929):619–624. doi: 10.1136/bmj.308.6929.619. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Matsumura Y., Sugiyama M., Matsumura S., Hayle A. J., Robinson P., Smith J. C., Tarin D. Unusual retention of introns in CD44 gene transcripts in bladder cancer provides new diagnostic and clinical oncological opportunities. J Pathol. 1995 Sep;177(1):11–20. doi: 10.1002/path.1711770104. [DOI] [PubMed] [Google Scholar]
- Matsumura Y., Tarin D. Significance of CD44 gene products for cancer diagnosis and disease evaluation. Lancet. 1992 Oct 31;340(8827):1053–1058. doi: 10.1016/0140-6736(92)93077-z. [DOI] [PubMed] [Google Scholar]
- Shibata D., Almoguera C., Forrester K., Dunitz J., Martin S. E., Cosgrove M. M., Perucho M., Arnheim N. Detection of c-K-ras mutations in fine needle aspirates from human pancreatic adenocarcinomas. Cancer Res. 1990 Feb 15;50(4):1279–1283. [PubMed] [Google Scholar]
- Shibata D., Cosgrove M., Arnheim N., Martin W. J., Martin S. E. Detection of human papillomavirus DNA in fine-needle aspirations of metastatic squamous-cell carcinoma of the uterine cervix using the polymerase chain reaction. Diagn Cytopathol. 1989;5(1):40–43. doi: 10.1002/dc.2840050109. [DOI] [PubMed] [Google Scholar]


