A new approach to epithelial–mesenchymal transition diagnostics in epithelial tumors: double immunofluorescent staining and flow cytometry
Tatiana A Bogush,
Anna A Basharina,
Baina K Eliseeva,
Sergey A Kaliuzhny,
Elena A Bogush,
Vladislav Y Kirsanov,
Mikhail M Davydov,
Vyacheslav S Kosorukov
Affiliations
Tatiana A Bogush
1Department of Tumor Molecular Markers, N.N. Blokhin National Medical Research Center of Oncology of the Ministry of Health of the Russian Federation, 24 Kashirskoe Shosse, Moscow, Russia
Anna A Basharina
1Department of Tumor Molecular Markers, N.N. Blokhin National Medical Research Center of Oncology of the Ministry of Health of the Russian Federation, 24 Kashirskoe Shosse, Moscow, Russia
Baina K Eliseeva
1Department of Tumor Molecular Markers, N.N. Blokhin National Medical Research Center of Oncology of the Ministry of Health of the Russian Federation, 24 Kashirskoe Shosse, Moscow, Russia
Sergey A Kaliuzhny
1Department of Tumor Molecular Markers, N.N. Blokhin National Medical Research Center of Oncology of the Ministry of Health of the Russian Federation, 24 Kashirskoe Shosse, Moscow, Russia
Elena A Bogush
2Oncological Department of Surgery no. 2, N.N. Blokhin National Medical Research Center of Oncology of the Ministry of Health of the Russian Federation, 23 Kashirskoe Shosse, Moscow, Russia
Vladislav Y Kirsanov
3Department of Oncology, N.V. Sklifosovsky Institute of Clinical Medicine, I.M. Sechenov First Moscow State Medical University of the Ministry of Health of the Russian Federation (Sechenov University), 8-2 Trubetskaya str., Moscow, Russia
Mikhail M Davydov
3Department of Oncology, N.V. Sklifosovsky Institute of Clinical Medicine, I.M. Sechenov First Moscow State Medical University of the Ministry of Health of the Russian Federation (Sechenov University), 8-2 Trubetskaya str., Moscow, Russia
Vyacheslav S Kosorukov
4Laboratory of Transgenic Drugs, N.N. Blokhin National Medical Research Center of Oncology of the Ministry of Health of the Russian Federation, 24 Kashirskoe Shosse, Moscow, Russia
A new method of double immunofluorescent staining for flow cytometry has been created to evaluate quantitative expression of mesenchymal protein vimentin only in epithelial cells of a solid tumor that is a mix of different origin cells. De novo vimentin expression is strongly associated with epithelial–mesenchymal transition and therefore is a metastatic potential marker of epithelial tumor cells. In comparison with semiquantitative available methods, the proposed one has several advantages, such as the accurate measurement of the marker’s expression, and minimization of spatial and temporal tumor heterogeneity. Clinical validation of the method has revealed inverse correlation between the quantitative index of epithelial–mesenchymal transition level and progression-free survival using Kaplan–Meier curves and the COX proportional hazards ratio in 32 ovarian cancer patients.
