MODELING OF A SUBCUTANEOUS XENOGRAFT OF HUMAN SKIN MELANOMA MEL CHER WITH V600E BRAF MUTATION IN IMMUNODEFICIENT MICE FOR PRECLINICAL STUDY THE TARGETING ANTICANCER DRUGS

Introduction. Development of new models of a human disseminated skin melanoma of the with molecular-genetic targets for specific therapy increases productivity of the preclinical researches new the anti-melanoma drugs or their combinations in vitro and in vivo. Such opportunity is realized by adaptation in vivo of the original human pigmented skin melanoma cell line mel Cher and receiving subcutaneous (s. c.) xenograft under monitoring of transplant, morphological, molecular-genetic (V600E BRAF mutation) and chemotherapeutic (sensitivity for the inhibitor of BRAF kinases to a vemurafenib) characteristics. Objective: receiving from the cell line mel Cher s. c. xenogratft of the human pigmented skin melanoma with V600E BRAF mutation and sensitive to specific target therapy. Materials and methods. Human pigmented skin melanoma cell line mel Cher from the Collection of Russian Cancer Research Center and immunodeficient Balb/c nude female mice cultivated in Russian Cancer Research Center was used. Required characteristics are defined by multiple s. c. transplanting in vivo by methods of transplant biology, a light microscopy, molecular-genetics and the experimental chemotherapy. Sensitivity to a BRAF kinase inhibitor to a vemurafenib was estimated under monitoring of the tumor growth rate (Vt/V0) on indexes, adequate for patients: existence of the complete remission and possibility of recurrence. Results. When s. c. transplantation of 107 cell of mel Cher line cytological identical intertwined s. c. xenografts with a stable growth kinetics on 4-9 passages (a latent phase 8 days, exponential - to 14 days, stationary - to 24 days) and existence of a mutation of V600E BRAF have been recieved. Vemurafenib in a single dose of 75 mg/kg caused the complete remission during a 15-day course and within 7 days after its cancellation - with the subsequent recurrence. Conclusion: receiving from the cell line mel Cher s. c. xenogratft of a human pigmented melanoma of skin with a mutation of V600E BRAF and sensitive to specific target therapy is suitable for preclinical studying of the new anti-melanoma drugs specific for this target.

пигментированная меланома кожи человека, мутация V600E BRAF, подкожный ксенографт, иммунодефицитная мышь, human pigmented skin melanoma, V600E BRAF mutation, subcutaneous xenograft, immunodeficient mice

1. Вишневская Я.В., Машенкина Я.А., Сендерович А.И. и др. Озвременная морфологическая, иммуногистохимическая и молекулярно-генетическая диагностика меланомы кожи. Сибирский онкологический журнал 2012;4(52): 74-5.

2. Демидов Л.В., Орлова К.В. Индивидуализация лекарственного лечения меланомы кожи. Практическая онкология 2013;14(4):239-46.

3. Bollag G., Hirth P., Tsai J. et al. Clinical efficacy of a RAF inhibitor needs broad target blockade in BRAF-mutant melanoma. Nature 2010;467:596-9.

4. Flaherty K.T., Puzanov I., Kim K.B. et al. Inhibition of mutated, activated BRAF in metastatic melanoma. N Engl J Med 2010;363:809-19.

5. Joseph E.W., Pratilas C.A., Poulikakos P.I. et al. The RAF inhibitor PLX4032 inhibits ERK signaling and tumor cell proliferation in a V600E BRAF-selective manner. Proc Natl Acad Sci U S A 2010;107(33):14903-8.

6. Tsai J., Lee J.T., Wang W. et al. Discovery of a selective inhibitor of oncogenic B-Raf kinase with potent antimelanoma activity. Proc Natl Acad Sci USA 2008; 105:3041-6.

7. Benlloch S., Paya A., Alenda C. et al. Detection of BRAF V600E mutation in colorectal cancer comparison of automatic sequencing and real-time chemistry methodology. J Mol Diagn 2006;8:540-3.

8. Ribas A., Kim K.B., Schuchter L.M. et al. BRIM-2: an open label, multicenter phase II study of vemurafenib in previously treated patients with BRAF V600E mutation positive melanoma. J Clin Oncol 2011; 29(15 Suppl):8509.

9. Walker G.J., Soyer H.P., Terzian T., Box N.F. Modelling melanoma in mice. Pigment Cell Melanoma Res 2011;24:1158-76.

10. Михайлова И.М., Барышников А.Ю. Коллекция клеточных линий меланомы человека. Издательство НИИ ЭДиТО ФГБУ «РОНЦ им. Н.Н. Блохина» Минздрава России 2016:107.

11. Патент РФ № 2364624. Клеточная линия меланомы человека mel Cher, используемая для получения противоопухолевых вакцин. Михайлова И.Н., Барышников А.Ю., Демидов Л.В., Киселев С.Л., Бурова О.С., Морозова Л.Ф. 2009.

12. Трещалина Е.М. Иммунодефицитные мыши разведения РОНЦ им. Н.Н. Блохина РАМН. Возможности использования. М., 2010.

13. Патент РФ № 2572569. Способ получения подкожных ксенографтов клеточной линии меланомы кожи человека mel Cher с мутацией V600E BRAF для доклинического изучения таргетных противоопухолевых средств. 2015.

14. Трещалина Е.М., Андронова Н.В., Гарин А.М. Доклиническое изучение противоопухолевых препаратов. В кн.: Рациональная фармакотерапия в онко логии. М.: Litterra, 2015. С. 75-82.