Detection of melanoma cells in peripheral blood by means of reverse transcriptase and polymerase chain reaction

doi:10.1016/0140-6736(91)92100-G

The Lancet

Volume 338, Issue 8777, 16 November 1991, Pages 1227-1229

https://doi.org/10.1016/0140-6736(91)92100-G Get rights and content

Abstract

Only small numbers of cells from solid tumours are needed for haematogenous metastasis. Detection is difficult because existing techniques are not sensitive enough. We have used reverse transcriptase to make complementary DNA from peripheral blood messenger RNA, and the polymerase chain reaction (PCR) to amplify cDNA specific for a gene actively transcribed only in the tumour tissue type. We prepared c DNA from peripheral blood of seven patients with malignant melanoma, four patients with other metastatic cancers, and four healthy subjects, as well as from several melanoma-derived cell lines. PCR was used to amplify the gene for tyrosinase, a tissue-specific gene in melanocytes. Since normal melanocytes are not thought to circulate in peripheral blood, detection of tyrosinase transcription in peripheral blood should indicate the presence of circulating cancer cells. The method was highly sensitive and could detect a single melanoma cell from a cell line in 2 ml normal blood. Blood samples from four of the seven patients with malignant melanoma gave positive results, whereas all eight control subjects gave negative results. This method does not depend on the characterisation of cancer-specific genetic abnormalities and can be applied to any cancer for which tissue-specific genes can be identified, including epithelial cancers. It could prove useful in the diagnosis of primary or metastatic cancers, in assessing prognosis, and in detecting residual disease after treatment.

References (16)

J. Lyons
Analysis of ras gene point mutations by PCR and oligonucleotide hybridization
Y. Tomita et al.
Human oculocutaneous albinism caused by single base insertion in the tyrosinase gene
Biochem Biophys Res Commun
(1989)
Rm Lawn et al.
The nucleotide sequence of the human β-globin gene
Cell
(1980)
H. Naito et al.
Detection of tyrosine hydroxylase mRNA and minimal neuroblastoma cells by the reverse transcription-polymerase chain reaction
Eur J Cancer
(1991)
Y-MD Lo et al.
Prenatal sex determination by DNA amplification from maternal peripheral blood
Lancet
(1989)
D. Tarin et al.
Clinicopathological observations on metastases in man studied in patients treated with peritoneovenous shunts
Br Med J
(1984)
Tj Moss et al.
Detection of neuroblastoma cells in blood
J Clin Oncol
(1990)

There are more references available in the full text version of this article.

Cited by (633)

An electrochemical biosensor based on network-like DNA nanoprobes for detection of mesenchymal circulating tumor cells
2023, Biosensors and Bioelectronics
The identification and detection of mesenchymal circulating tumor cells （mCTCs） is important for early warning of tumor metastasis. The majority of conventional detection methods for CTCs rely on the recognition of epithelial biomarkers, which is technically challenging for detecting CTCs with epithelial-mesenchymal transition (EMT)-induced phenotypic alteration. In this work, we have constructed a label-free biosensor for sensitive electrochemical assay of mCTCs. In our design, the capture probe can recognize the vimentin overexpressed on the surface of mCTCs with high specificity. Meantime, the network-like DNA nanoprobes with multiple G-quadruplex/hemin complexes and multiple cholesterol molecules can be grafted to the cell surface based on the high affinity between cholesterol molecules and cell membrane. Owing to the mimic horseradish peroxidase of G-quadruplex/hemin complexes, strong electrochemical responses will be obtained for sensitive quantification of mCTCs with a detection limit of 8 cell mL⁻¹. Moreover, the biosensor can effectively overcome the interference of vimentin negative cells or secretory vimentin, and realize the recovery tests in whole blood with high accuracy, thereby may further promoting the diagnosis and personalized treatment of cancer in clinic.
Circulating Plasma Tumor DNA Is Superior to Plasma Tumor RNA Detection in Ewing Sarcoma Patients: ptDNA and ptRNA in Ewing Sarcoma
2021, Journal of Molecular Diagnostics
The detection of tumor-specific nucleic acids from blood increasingly is being used as a method of liquid biopsy and minimal residual disease detection. However, achieving high sensitivity and high specificity remains a challenge. Here, we perform a direct comparison of two droplet digital PCR (ddPCR)-based detection methods, circulating plasma tumor RNA and circulating plasma tumor DNA (ptDNA), in blood samples from newly diagnosed Ewing sarcoma patients. First, we developed three specific ddPCR-based assays to detect EWS-FLI1 or EWS-ERG fusion transcripts, which naturally showed superior sensitivity to DNA detection on in vitro control samples. Next, we identified the patient-specific EWS-FLI1 or EWS-ERG breakpoint from five patient tumor samples and designed ddPCR-based, patient-specific ptDNA assays for each patient. These patient-specific assays show that although plasma tumor RNA can be detected in select newly diagnosed patients, positive results are low and statistically unreliable compared with ptDNA assays, which reproducibly detect robust positive results across most patients. Furthermore, the unique disease biology of Ewing sarcoma enabled us to show that most cell-free RNA is not tumor-derived, although cell-free-DNA burden is affected strongly by tumor-derived DNA burden. Here, we conclude that, even with optimized highly sensitive and specific assays, tumor DNA detection is superior to RNA detection in Ewing sarcoma patients.
Single-Cell Diagnostics, Prognosis, and Therapy
2019, Single-Cell Omics
Single-cell diagnostics, prognosis, and therapy
2019, Single-Cell Omics: Volume 2: Application in Biomedicine and Agriculture
An Update Regarding the Molecular Genetics of Melanocytic Neoplasms and the Current Applications of Molecular Genetic Technologies in Their Diagnosis and Treatment
2018, Clinics in Laboratory Medicine
Circulating tumor cells count as a predictor of survival in lung cancer
2018, Critical Reviews in Oncology/Hematology
Citation Excerpt :
Conventional or automated scanning microscopes and cytometers, in combination with immunocytochemistry (ICC) or immunofluorescence for the expression of various epithelial (e.g. cytokeratins), mesenchymal or tissue-specific markers −along with staining for the nuclear dye 4′, 6-doamidino-2-phenylindole (DAPI)- allow the detection and enumeration of CTCs (Joosse et al., 2015; Krebs et al., 2010; Toss et al., 2014; Millner et al., 2013). Nucleic acid-based assays target gene alterations or tumor-specific mRNA transcripts, mainly employing qualitative or quantitative reverse transcriptase-PCR (RT-PCR or qRT-PCR), while functional assays, like the EPithelial Immuno SPOT technology (EPISPOT) detect cell-secreted proteins, thus leading to isolation of only viable CTCs (Toss et al., 2014; Smith et al., 1991; Stathopoulou et al., 2003; Alix-Panabières and Pantel, 2015; Soler et al., 2017). Assays for CTC isolation and analysis, combining enrichment and detection steps (e.g. CellSearch, AdnaTest, and the above described techniques ISET and EPISPOT), are also commercially available (Vona et al., 2000; Soler et al., 2017; Allard et al., 2004; Allard and Terstappen, 2015; Andreopoulou et al., 2012).
The presence of circulating tumor cells (CTCs) in the peripheral blood of cancer patients was first described in the second half of the 19th century, but research interest in their potential clinical utility has intensified and greatly expanded only in recent years. Herein, we summarize and critically discuss current knowledge on CTC count as a predictor of survival in lung cancer, and comment on the existing challenges and future perspectives in this field. The majority of data published to date, including the results of almost all large cohorts, are strongly supportive of the value of CTC enumeration as a predictor of survival, mainly in advanced/metastatic non-small and small cell lung cancer (NSCLC and SCLC, respectively). Nonetheless, additional research is warranted to establish the prognostic relevance of CTC count in other clinical settings, mainly encompassing earlier-stage disease as well as specific molecular subtypes of NSCLC (e.g. EGFR mutation-positive or ALK-positive cases).

View all citing articles on Scopus

View full text

Detection of melanoma cells in peripheral blood by means of reverse transcriptase and polymerase chain reaction

Abstract

Biochem Biophys Res Commun

Cell

Eur J Cancer

Lancet

Clinicopathological observations on metastases in man studied in patients treated with peritoneovenous shunts

Br Med J

Detection of neuroblastoma cells in blood

J Clin Oncol