MODELING OF ORTHOTOPICAL GROWTH OF HUMAN KIDNEY CANCER WITH SUBCUTANEOUS XENOGRAFTS RPOCH1 ON IMMUNODEFICIENT MICE

Introduction. Creation of new orthotopic models of tumor growth (surgical orthotopic implantation, SOI) promotes pre-clinical studies of anti-metastatic functional potential of the drugs including those which aim to treat advanced clear-cell renal carcinoma with the use of patient’s tumor or metastasis sample for implantation material. The use of subcutaneous (s. c.) xenograft contributes to SOI improvement and extends experimental opportunities. The collection of tumor strains of N.N. Blokhin Russian Cancer Research Center that includes s. c. xenograft RPoch1, which was characterized by transplantation, pathological, and immunological parameters and in terms of sensitivity towards classical antitumor cytostatics, presents the base for obtaining an orthotopic model for multifunctional usage in pre-clinical studies of specific medicines. Objective. Preparation and obtaining an orthotopic model of human clear-cell renal carcinoma with the implant of subcutaneous xenograft RPochl. Materials and methods. The study used s. c. RPoch1 and immunodeficient female mice Balb/c nude on the N. N. Blokhin Russian Cancer Research Center breeding. Transplantation characteristics were defined during implant passages into kidney parenchyma. The effectiveness of orthotopic implantation was estimated in the dynamics according to kinetic, pathological, morphological, and immunological parameters by light microscopy, flow cytometry, monoclonal antibodies (mAb, Caltag Laboratories) against corresponding antigens. Sensitivity to specific drugs was assessed against inhibitors of tyrosine kinases according to standard parameters (T/C <42 %, total regressions). Results. After orthotopic implantation of RPochl on days 7, 14 and 21 the tumor lesions were registered with the size of 5.0 ± 1.0 mm³, 215.0 ± 88.0 mm³ and 608.0 ± 157.0 mm³. Histological examination showed that the tumor had separated connective tissue layers of solid areas of large polymorphic cells with mitosis (10-15 in a lesion) and single areas with necrosis on day 21. The 1.5-fold enhanced expression of NK1.1 in mice on day 14 and 3.3-fold increase of CD8a expression demonstrate the tendency to develop immune suppression induced by growing tumor. The results showed high sensitivity of RPoch1 to sorafenib and sunitinib (T/C_mn = 2-30 %). Conclusion. The authors obtained an orthotopic xenograft of human clear-cell renal carcinoma RPoch1 with 100 % transplantation potential, reproducible biological characteristics including immunological parameters and sensitivity to inhibitors of tyrosine kinases, which can be used for pre-clinical studies of various agents including immunological or different combinations of antitumor drugs against this cancer type.

ортотопический имплантат, рак почки человека, orthotopic implant, human kidney cancer

1. Survey of antitumor and toxicity test systems. EORTC. Screening and Pharmacology Group 1989; Oct.: 23-25.

2. Трещалина Е.М., Жукова О.С., Герасимова Г.К. и др. Методические рекомендации по доклиническому изучению противоопухолевой активности лекарственных средств. В кн.: Руководство по проведению доклинических исследований лекарственных средств. Часть первая. М.: Гриф и К, 2012. С. 642-657.

3. Трещалина Е.М. Коллекция опухолевых штаммов человека. Под ред. М.И. Давыдова. М.: Практическая медицина, 2009. C. 37-38.

4. Самойленко И.В., Харкевич Г.Ю., Демидов Л.В. Применение блокатора рецепторов CTLA4 в лечении больных метастатической меланомой. Российский медицинский журнал 2015; 1: 4-9.

5. Носов Д.А. Рак почки. В кн.: Руководство по химиотерапии опухолевых заболеваний. Под ред. Н.И. Переводчиковой, В.А. Горбуновой. М.: Практическая медицина, 2015. С. 294-300.

6. Anticancer Drug Development Guide. Preclinical screening, clinical trials, and approval. Ed. by B.A. Teicher, P.A. Andrews. 2nd ed. Totowa. NJ: Humana Press, 2004. 450 p.

7. Michel S.M., Vervenn E.W., De Santis M. et al. SWITCH: A randomized sequential open-label study to evaluate efficacy safety of sorafenib (SO)/ sunitinib (SU) versus SU/SO in the treatment of metastatic renal cell cancer (mRCC). J Clin Oncol 2014; 32 (4): 393. DOI: 10.1016/j. eururo. 2015.04.017. PMID: 25952317.

8. Rini B.I., Escudier B., Tomczar P. et al. Comparative effectiveness of axitinib versus sorafenib in advanced renal cell carcinoma (AXIS): a randomised phase 3 trial. Lancet 2011; 378(9807): 1931-9. DOI: 10.1016/S0140-6736(11)61613-9. PMID: 22056247.

9. Трещалина Е.М. Иммунодефицитные мыши разведения РОНЦ им. Н.Н. Блохина РАМН. Возможности использования. М.: Издательская группа РОНЦ, 2010.

10. Шубина И.Ж., Чикилева И.О., Михайлова И.Н. и др. Активированные натуральные киллеры в клеточной иммунотерапии. Российский иммунологический журнал 2012; 6 (15); 1: 71-9.