Anti-pan Cytokeratin monoclonal antibody, clone 7F5&NR46

Circulating tumor cells (CTCs) are of recognized importance for diagnosis and prognosis of cancer patients. With melanoma, most studies do not show any clear relationship between CTC levels and stage of disease. Here, CTCs were enriched (similar to 400X) from blood of melanoma patients using a simple centrifugation device (OncoQuick), and 4 melanocyte target RNAs (TYR, MLANA, MITF, and MIF) were quantified using QPCR. Approximately one-third of melanoma patients had elevated MIF and MLANA transcripts (p<0.0001 and p<0.001, respectively) compared with healthy controls. In contrast, healthy controls had uniformly higher levels of TYR and MITF than melanoma patients (p<0.0001). There was a marked shift of leukocytes into the CTC-enriched fractions (a 430% increase in RNA recovery, p<0.001), and no relationship between CTC levels and stage of disease was found. CTCs were captured on microfabricated filters and cultured. Captured melanoma CTCs were large cells, and consisted of 2 subpopulations, based on immunoreactivity. One subpopulation (similar to 50%) stained for both pan-cytokeratin (KRT) markers and the common leukocyte marker CD-45, whereas the second subpopulation stained for only KRT. Since similar cells are described in many cancers, we also examined blood from colorectal and pancreatic cancer patients. We observed analogous results, with most captured CTCs staining for both CD-45/KRT markers (and for the monocyte differentiation marker CD-14). Our results suggest that immature melanocyte-related cells (expressing TYR and MITF RNA) may circulate in healthy controls, although they are not readily detectable without considerable enrichment. Further, as early-stage melanomas develop, immature melanocyte migration into the blood is somehow curtailed, whereas a significant proportion of patients develop elevated CTC levels (based on MIF and MLANA RNAs). The nature of the captured CTCs is consistent with literature describing leukocyte/macrophage-tumor cell fusion hybrids, and their role in metastatic progression.

Intraoperative pathologic diagnosis for central nervous system (CNS) tumors is important to determine the neurosurgery procedure. But sometimes the differential diagnosis between glioma and lymphoma, or glioma and metastatic tumors is difficult for a pathologist during a short time, especially when the specimen is small or the frozen section has ice crystals. Immunohistochemistry (IHC) is a very useful method for diagnosis, but the traditional immunohistochemical method is time consuming and not suitable intraoperatively. In this study, we chose Cytokeratin-pan, GFAP, and LCA as three immunohistochemical indicators. Intraoperative IHC was done by Novodiax ihcDirect technology combined with Leica Bond auto-staining. Compared with the manual method recommended for the reagents (Novodiax ihcDirect), the results show that auto-staining has better stability and high reproducibility in coloration, which has broad prospects for future application.