title: Comparative assessment of lymph node micrometastasis in cervical, endometrial and vulvar cancer: insights on the real time qRT-PCR approach versus immunohistochemistry, employing dual molecular markers.
abstract: To address the value of qRT-PCR and IHC in accurately detecting lymph node micrometastasis in gynecological cancer, we performed a systematic approach, using a set of dual molecular tumor-specific markers such as cytokeratin 19 (CK19) and carbonic anhydrase 9 (CA9), in a series of 46 patients (19 with cervical cancer, 18 with endometrial cancer, and 9 with vulvar cancer). A total of 1281 lymph nodes were analyzed and 28 were found positive by histopathology. Following this documentation, 82 lymph nodes, 11 positive and 71 negative, were randomly selected and further analyzed both by IHC and qRT-PCR for CK19 and CA9 expression. All 11 (100%) expressed CK19 by IHC, while only 6 (54.5%) expressed CA9. On the contrary, all the histologically negative for micrometastases lymph nodes were also negative by IHC analysis for both markers. The comparative diagnostic efficacy of the two markers using qRT-PCR, however, disclosed that the analysis of the same aliquots of the 82 lymph nodes led to 100% specificity for the CK19 biomarker, while, in contrast, CA9 failed to recapitulate a similar pattern. These data suggest that qRT-PCR exhibits a better diagnostic accuracy compared to IHC, while CK19 displays a consistent pattern of detection compared to CA9.
authors: First Department of Obstetrics and Gynecology, University of Athens School of Medicine, Alexandra Hospital, 11528 Athens, Greece ; Laboratory of Biology, Department of Basic Medical Sciences, University of Athens School of Medicine, 11527 Athens, Greece ; Laboratory of Cell and Gene Therapy, Center of Basic Research II, Biomedical Research Foundation of the Academy of Athens, 11527 Athens, Greece.
authors: Kalliopi I
authors: KI
authors: Pappa
authors: First Department of Obstetrics and Gynecology, University of Athens School of Medicine, Alexandra Hospital, 11528 Athens, Greece.
authors: Alexandros
authors: A
authors: Rodolakis
authors: First Department of Obstetrics and Gynecology, University of Athens School of Medicine, Alexandra Hospital, 11528 Athens, Greece.
authors: Ioanna
authors: I
authors: Christodoulou
authors: Laboratory of Biology, Department of Basic Medical Sciences, University of Athens School of Medicine, 11527 Athens, Greece ; Laboratory of Cell and Gene Therapy, Center of Basic Research II, Biomedical Research Foundation of the Academy of Athens, 11527 Athens, Greece.
authors: Maria
authors: M
authors: Gazouli
authors: Department of Pathology, Alexandra Hospital, 11528 Athens, Greece.
authors: Sofia
authors: S
authors: Markaki
authors: First Department of Obstetrics and Gynecology, University of Athens School of Medicine, Alexandra Hospital, 11528 Athens, Greece.
authors: Aris
authors: A
authors: Antsaklis
authors: Laboratory of Biology, Department of Basic Medical Sciences, University of Athens School of Medicine, 11527 Athens, Greece ; Laboratory of Cell and Gene Therapy, Center of Basic Research II, Biomedical Research Foundation of the Academy of Athens, 11527 Athens, Greece.
authors: Nicholas P
authors: NP
authors: Anagnou
mesh_terms: D000951:Antigens, Neoplasm / Q000096:biosynthesis; D014408:Biomarkers, Tumor / Q000096:biosynthesis; D000071231:Carbonic Anhydrase IX; D002256:Carbonic Anhydrases / Q000096:biosynthesis; D016889:Endometrial Neoplasms / Q000235:genetics* / Q000473:pathology; D005260:Female; D006801:Humans; D053539:Keratin-19 / Q000096:biosynthesis; D008198:Lymph Nodes / Q000378:metabolism / Q000473:pathology; D061206:Neoplasm Micrometastasis / Q000235:genetics* / Q000473:pathology; D009367:Neoplasm Staging; D002583:Uterine Cervical Neoplasms / Q000235:genetics* / Q000473:pathology; D014846:Vulvar Neoplasms / Q000235:genetics* / Q000473:pathology
chemical_list: D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor; D053539:Keratin-19; C089226:CA9 protein, human; D000071231:Carbonic Anhydrase IX; D002256:Carbonic Anhydrases