Quantitative real-time RT-PCR in sentinel lymph nodes from melanoma patients.

Apmis ◽  
2008 ◽  
Vol 116 (3) ◽  
pp. 199-205 ◽  
Author(s):  
RIKKE RIBER-HANSEN ◽  
HELENE NORTVIG ABRAHAMSEN ◽  
BOE SANDAHL SORENSEN ◽  
STEPHEN JACQUES HAMILTON-DUTOIT ◽  
TORBEN STEINICHE
2009 ◽  
Vol 16 (1) ◽  
Author(s):  
Josep M. Hilari ◽  
Cristina Mangas ◽  
Liqiang Xi ◽  
Cristina Paradelo ◽  
Carlos Ferrándiz ◽  
...  

2002 ◽  
Vol 64 (2) ◽  
pp. 211-217 ◽  
Author(s):  
Yoichi MOROI ◽  
Chikage OBATA ◽  
Shohei FUJITA ◽  
Satoko SHIBATA ◽  
Sawako KONISHI ◽  
...  

2004 ◽  
Vol 22 (14_suppl) ◽  
pp. 7562-7562
Author(s):  
M. Gonzalez Cao ◽  
C. Badenas ◽  
J. Malvehy ◽  
R. Martí ◽  
T. Castel ◽  
...  

2003 ◽  
Vol 21 (2) ◽  
pp. 306-312 ◽  
Author(s):  
Angela Gradilone ◽  
Paola Gazzaniga ◽  
Diego Ribuffo ◽  
Susanna Scarpa ◽  
Emanuele Cigna ◽  
...  

Purpose: The expression of apoptosis-related genes, such as survivin, bcl-2, bcl-X, and bax, has been evaluated by reverse transcriptase polymerase chain reaction (RT-PCR) and by immunohistochemistry in sentinel lymph nodes (SLNs) from melanoma patients and then correlated to the outcome of patients. Patients and Methods: Thirty-six SLNs were examined. After RNA extraction, an RT-PCR followed by Southern blot hybridization was performed to detect survivin, bcl-2, bcl-X, and bax mRNA. bcl-2, survivin, and bax gene expression was evaluated, whenever possible, also by immunohistochemistry at the protein level. Results: We found a significant correlation (P < .005) between survivin expression and outcome of patients; in fact, 61.5% of patients expressing survivin gene progressed or died because of the disease, whereas 38.5% are currently disease-free. Among patients negative for survivin expression, 100% are disease-free after a median follow-up time of 52.9 months. We did not find a significant correlation between bcl-2, bax, and bcl-X gene expression and outcome of patients. In fact, these genes were found equally expressed in patients with disease progression and in disease-free patients. Conclusion: Our findings show a variable expression of apoptosis-related genes in SLNs of melanoma patients; more interestingly, we found that survivin expression correlates to outcome of patients in a statistically significant way, whereas the expression of other genes, such as bcl-2, bax, and bcl-X, did not seem to correlate to progression of disease. We suggest that the detection of survivin gene expression by RT-PCR in SLNs may be a useful prognostic indicator.


2005 ◽  
Vol 124 (3) ◽  
pp. 633-637 ◽  
Author(s):  
Thomas Giese ◽  
Monika Engstner ◽  
Ulrich Mansmann ◽  
Wolfgang Hartschuh ◽  
Bernhard Arden

2004 ◽  
Vol 22 (14_suppl) ◽  
pp. 7562-7562
Author(s):  
M. Gonzalez Cao ◽  
C. Badenas ◽  
J. Malvehy ◽  
R. Martí ◽  
T. Castel ◽  
...  

2009 ◽  
Vol 27 (15_suppl) ◽  
pp. 628-628
Author(s):  
M. Kurosumi ◽  
Y. Kobayashi ◽  
H. Takei

628 Background: Analysis using real time RT-PCR for the detection of metastases in lymph nodes (LN) increases sampling but is associated with a risk of too much sensitivity as compared to histological analysis. Determining the appropriate cutoff value is important in introducing this system to routine clinical practice. Our study confirms the reliability of the cutoff values of a real-time RT-PCR assay (GeneSearch, Veridex LLC) to detect metastases larger than 0.2 mm by detailed 0.2 mm frozen section histological diagnosis. Methods: 129 sentinel and non-sentinel lymph nodes were obtained by sentinel LN biopsy from 79 breast cancer patients. All LNs were cut in half. One half of each LN was used for routine intra-operative diagnosis. The other LN half had sections cut every 0.2 mm for histological analysis. All frozen sections were H&E stained and examined by on-site pathologists. These sections were then re-stained immunohistochemically using a pancytokeratin antibody (AE1/AE3) for detecting submicrometastses. All remaining tissue between the histology sections was assayed by the real-time RT-PCR assay using the genetic markers mammaglobin (MG) and cytokeratin-19 (CK-19) (note: tissue processing method was investigational and off-label). Cutoff values were pre-set in a large US study (n = 304). Results: Compared to the histological diagnosis using 0.2 mm interval frozen sections, the real-time RT-PCR results were as follows; sensitivity of 100.0% (34/34), specificity of 93.7% (89/95), and overall accuracy of 95.3% (123/129). Submicrometastases were recognized by IHC in 3 of 95 samples (2 solitary and 1 multiple sites) diagnosed as negative by H&E-stain sections, and 3 samples were negative for RT-PCR. Conclusions: Results of 0.2 mm interval histological analysis suggest a near perfect sensitivity of the real time RT-PCR assay, allowing reliable detection of LN metastases larger than 0.2 mm. In addition, at least three samples with submicrometastases detected by immunohistochemistry for pancytokeratin were above the preset cutoff values of this real time RT-PCR assay. [Table: see text]


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