Impact of siRNA transfection on tumor development under experimental pathology of thyroid gland

Aim. To assess the impact of siRNA transfection on the tumor development under experimental pathology of the thyroid gland. Methods. Experiments were performed on rats weighing 180 ± 20 g, which were divided into five groups: IA and IB – animals with simulated hypo- and hyperthyroid states and trans...

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Дата:	2013
Автори:	Zaporozhan, V.M., Maryniuk, G.S., Kholodkova, O.L., Bubnov, V.V., Andronov, D.U.
Формат:	Стаття
Мова:	English
Опубліковано:	Інститут молекулярної біології і генетики НАН України 2013
Назва видання:	Вiopolymers and Cell
Теми:	Structure and Function of Biopolymers
Онлайн доступ:	http://dspace.nbuv.gov.ua/handle/123456789/153246
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Назва журналу:	Digital Library of Periodicals of National Academy of Sciences of Ukraine
Цитувати:	Impact of siRNA transfection on tumor development under experimental pathology of thyroid gland / V.M. Zaporozhan, G.S. Maryniuk, O.L. Kholodkova, V.V. Bubnov, D.U. Andronov // Вiopolymers and Cell. — 2013. — Т. 29, №. 6. — С. 463-467. — Бібліогр.: 16 назв. — англ.

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spelling	irk-123456789-1532462019-06-14T01:28:19Z Impact of siRNA transfection on tumor development under experimental pathology of thyroid gland Zaporozhan, V.M. Maryniuk, G.S. Kholodkova, O.L. Bubnov, V.V. Andronov, D.U. Structure and Function of Biopolymers Aim. To assess the impact of siRNA transfection on the tumor development under experimental pathology of the thyroid gland. Methods. Experiments were performed on rats weighing 180 ± 20 g, which were divided into five groups: IA and IB – animals with simulated hypo- and hyperthyroid states and transplanted Guerin’s carcinoma; IIA and IIB – animals with simulated hypo- and hyperthyroid states and transplanted Guerin’s carcinoma in combination with siRNA transfection; III – control group with transplanted Guerin’s carcinoma in combination with siRNA transfection. Orthogonal dimensions of the tumor were measured. Histological and immunohistochemical researches of tumor samples were performed. Results. It has been shown that the inhibitory effect of short interfering RNA is realized to a greater extent in the hypothyroid state, indicating an important role of thyroid hormones in the regulation of expression of genes that control the cell cycle. We discuss a possible dependence of necrobiotic processes prevalence and characteristic changes in the blood vessels on the effects of thyroid hormones on angiogenesis and proliferative processes. Conclusions. siRNA trasfection leads to inhibition of tumor growth in animals with both hypo- and hyperthyroidism, but it is more pronounced at hypothyroidism. Мета. Оцінити вплив трансфекційно введеної siРНК на розвиток пухлини за умов експериментальної патології щитоподібної залози. Методи. Експерименти виконували на самицях щурів масою 180 ± 20 г. Тварин розділено на п’ять груп: IA і IБ – щури з модельованими гіпо- і гіпертиреозом та перевитою карциномою Герена, IIA і IIБ – з модельованими гіпо- і гіпертиреозом та перевитою карциномою Герена у комплексі з трансфекційно введеною siРНК, ІІІ – з перевитою карциномою Герена у комплексі з трансфекційно введеною siРНК. Вимірювали ортогональні розміри пухлини. Проводили гістологічні та імуногістохімічні дослідження зразків пухлини. Результати. Показано, що інгібувальний вплив коротких інтерферуючих РНК більшою мірою проявляється за гіпотиреоїдного стану, що свідчить про важливу роль гормонів щитоподібної залози у регуляції експресії генів, контролюючих клітинний цикл. Обговорюється ймовірність залежності переважання некробіотичних процесів та характерних змін у судинах від дії тиреоїдних гормонів на ангіогенез і проліферативні процеси. Висновки. Трасфекція siРНК призводить до пригнічення пухлинного росту у тварин як з гіпо-, так і з гіпертиреозом, проте активніше цей процес протікає при гіпотиреоїдному стані. Цель. Оценка влияния трансфекционно введенной siРНК на развитие опухоли в условиях экспериментальной патологии щитовидной железы. Методы. Эксперименты проводили на самках крыс массой 180 ± 20 г. Животных разделили на пять групп: IA и IБ – крысы с моделированными гипо- и гипертиреозом и перевитой карциномой Герена, IIA и IIБ – с моделированными гипо- и гипертиреозом и перевитой карциномой Герена в комплексе с трансфекционно введенной siРНК, III – контрольная группа с перевитой карциномой Герена в комплексе с трансфекционно введенной siРНК. Измеряли ортогональные размеры опухоли. Осуществляли гистологические и иммуногистохимические исследования образцов опухоли. Результаты. Показано, что ингибирующее влияние коротких интерферирующих РНК в большей степени проявляется при гипотиреоидном состояния, что указывает на важную роль гормонов щитовидной железы в регуляции экспрессии генов, контролирующих клеточный цикл. Обсуждается вероятность зависимости преобладания некробиотических процессов и характерных изменений в сосудах от влияния тиреоидных гормонов на ангиогенез и пролиферативные процессы. Выводы. Трансфекция siРНК приводит к угнетению опухолевого роста у животных как с гипо-, так и с гипертиреозом, однако более активно этот процесс протекает при гипотиреоидного состоянии. 2013 Article Impact of siRNA transfection on tumor development under experimental pathology of thyroid gland / V.M. Zaporozhan, G.S. Maryniuk, O.L. Kholodkova, V.V. Bubnov, D.U. Andronov // Вiopolymers and Cell. — 2013. — Т. 29, №. 6. — С. 463-467. — Бібліогр.: 16 назв. — англ. 0233-7657 DOI: http://dx.doi.org/10.7124/bc.00083A http://dspace.nbuv.gov.ua/handle/123456789/153246 616.441-008.63:618.145-006.6:612-092.9 en Вiopolymers and Cell Інститут молекулярної біології і генетики НАН України
institution	Digital Library of Periodicals of National Academy of Sciences of Ukraine
collection	DSpace DC
language	English
topic	Structure and Function of Biopolymers Structure and Function of Biopolymers
spellingShingle	Structure and Function of Biopolymers Structure and Function of Biopolymers Zaporozhan, V.M. Maryniuk, G.S. Kholodkova, O.L. Bubnov, V.V. Andronov, D.U. Impact of siRNA transfection on tumor development under experimental pathology of thyroid gland Вiopolymers and Cell
description	Aim. To assess the impact of siRNA transfection on the tumor development under experimental pathology of the thyroid gland. Methods. Experiments were performed on rats weighing 180 ± 20 g, which were divided into five groups: IA and IB – animals with simulated hypo- and hyperthyroid states and transplanted Guerin’s carcinoma; IIA and IIB – animals with simulated hypo- and hyperthyroid states and transplanted Guerin’s carcinoma in combination with siRNA transfection; III – control group with transplanted Guerin’s carcinoma in combination with siRNA transfection. Orthogonal dimensions of the tumor were measured. Histological and immunohistochemical researches of tumor samples were performed. Results. It has been shown that the inhibitory effect of short interfering RNA is realized to a greater extent in the hypothyroid state, indicating an important role of thyroid hormones in the regulation of expression of genes that control the cell cycle. We discuss a possible dependence of necrobiotic processes prevalence and characteristic changes in the blood vessels on the effects of thyroid hormones on angiogenesis and proliferative processes. Conclusions. siRNA trasfection leads to inhibition of tumor growth in animals with both hypo- and hyperthyroidism, but it is more pronounced at hypothyroidism.
format	Article
author	Zaporozhan, V.M. Maryniuk, G.S. Kholodkova, O.L. Bubnov, V.V. Andronov, D.U.
author_facet	Zaporozhan, V.M. Maryniuk, G.S. Kholodkova, O.L. Bubnov, V.V. Andronov, D.U.
author_sort	Zaporozhan, V.M.
title	Impact of siRNA transfection on tumor development under experimental pathology of thyroid gland
title_short	Impact of siRNA transfection on tumor development under experimental pathology of thyroid gland
title_full	Impact of siRNA transfection on tumor development under experimental pathology of thyroid gland
title_fullStr	Impact of siRNA transfection on tumor development under experimental pathology of thyroid gland
title_full_unstemmed	Impact of siRNA transfection on tumor development under experimental pathology of thyroid gland
title_sort	impact of sirna transfection on tumor development under experimental pathology of thyroid gland
publisher	Інститут молекулярної біології і генетики НАН України
publishDate	2013
topic_facet	Structure and Function of Biopolymers
url	http://dspace.nbuv.gov.ua/handle/123456789/153246
citation_txt	Impact of siRNA transfection on tumor development under experimental pathology of thyroid gland / V.M. Zaporozhan, G.S. Maryniuk, O.L. Kholodkova, V.V. Bubnov, D.U. Andronov // Вiopolymers and Cell. — 2013. — Т. 29, №. 6. — С. 463-467. — Бібліогр.: 16 назв. — англ.
series	Вiopolymers and Cell
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first_indexed	2025-07-14T04:53:16Z
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fulltext	UDC 616.441-008.63:618.145-006.6:612-092.9 Impact of siRNA transfection on tumor development under experimental pathology of thyroid gland V. M. Zaporozhan, G. S. Maryniuk, O. L. Kholodkova, V. V. Bubnov, D. U. Andronov The Odessa National Medical University, Ukraine 2, Valihovskyy Str., Odessa, Ukraine, 65082 a_borshchenko@ukr.net; kholodkova_l@rambler.ru Aim. To assess the impact of siRNA transfection on the tumor development under experimental pathology of the thyroid gland. Methods. Experiments were performed on rats weighing 180 ± 20 g, which were divided into five groups: IA and IB – animals with simulated hypo- and hyperthyroid states and transplanted Guerin’s carcinoma; IIA and IIB – animals with simulated hypo- and hyperthyroid states and transplanted Guerin’s carcinoma in combination with siRNA transfection; III – control group with transplanted Guerin’s carcinoma in combination with siRNA transfection. Orthogonal dimensions of the tumor were measured. Histological and im- munohistochemical researches of tumor samples were performed. Results. It has been shown that the inhibitory effect of short interfering RNA is realized to a greater extent in the hypothyroid state, indicating an important role of thyroid hormones in the regulation of expression of genes that control the cell cycle. We discuss a possible dependence of necrobiotic processes prevalence and characteristic changes in the blood vessels on the effects of thyroid hormones on angiogenesis and proliferative processes. Conclusions. siRNA trasfection leads to in- hibition of tumor growth in animals with both hypo- and hyperthyroidism, but it is more pronounced at hypothyroidism. Keywords: transfection, siRNA, experimental model, hypothyroidism, hyperthyroidism. Introduction. Some authors consider the influence of T-helper subpopulations and other immune cells to be the basic mechanisms of the thyroid hormone (TH) ac- tion along with direct stimulation of cancerogenesis [1]. Thyroxine regulates the functioning of tyrosine protein and plays a significant role in apoptosis [2]. Thus, the functional state of the thyroid gland is an important pre- dictor of clinical outcome of tumors. There has been recently detected a possibility of the gene expression inhibition based on the creation of double-stranded RNA (dsRNA) molecules. Using the- se dsRNA enables target exclusion of individual genes without violation of the formation of neighboring ge- nes’ proteins; this method being more effective and fast than any other. This phenomenon is called RNA interfe- rention; the dsRNA-sequence that contributes to it is called siRNA (short interfering RNA). siRNA does not prevent gene readouts but includes the typical cell mechanism blocking the transcription of gene mRNA and prevents the formation of a corres- ponding protein (post-transcriptional gene signalling). Nonspecific siRNA binding with genes is an im- portant issue for researchers and clinicians who use them for gene expression inhibition. However, no pro- per attention has been paid to the application of tumor growth epigenetic modifications with thyroid dysfunc- tion in experimental models of low-differentiated tu- mors [3–5]. The SOCS family proteins form a]part of the classi- cal negative feedback system that regulates cytokine signal transduction. SOCS1 is involved in negative re- gulation of cytokines that signal through the JAK/ 463 ISSN 0233–7657. Biopolymers and Cell. 2013. Vol. 29. N 6. P. 463–467 doi: 10.7124/bc.00083A � Institute of Molecular Biology and Genetics, NAS of Ukraine, 2013 STAT3 pathway. Through binding to JAKs, SOCS1 in- hibits their kinase activity and suppresses the Tec pro- tein-tyrosine activity in vitro. There are known some siRNA modulating the SOCS1 gene expression, e. g. hsa-miR-411, hsa-miR-142-5p, hsa-miR-30c, hsa-miR- 379, hsa-miR-331-3p, hsa-miR-548v, hsa-miR-30d, hsa-miR-221, however their effects on the dynamics of tumor growth in the conditions of disorders of thyroid gland functions have been never investigated. The aim of the study was to assess an impact of siRNA transfection on the tumor development under ex- perimental pathology of the thyroid gland. Materials and methods. Experiments were conduc- ted on 32 white female laboratory rats weighing 180 ± � 20 g, which were kept on a vivarium standard diet. Guerin’s carcinoma (0.5 ml suspension of tumor cells – (5–6) �106 cells/ml) was transplanted to the animals of I group (n = 12) by subcutaneous injection in the back between the shoulders. These cells were obtained from donor rats (tumor strain was provided by R. E. Kavetsky Institute of Experimental Pathology, Oncology and Ra- diobiology, NAS of Ukraine). To the animals of II group (n = 12) the tumor cells were administered along with siRNA transfection according to the same method [6]. Each group was divided into two subgroups (A and B) depending on the functional state of thyroid gland. The hypothyroid state (subgroups IA and IIA) was si- mulated by blocking the thyroid hormones secretion by mercazolil that was administered one time per day at a dose of 5 mg per 100 g body weight intragastric within two weeks. The hyperthyroid condition (subgroups IB and IIB) was modeled by the L-thyroxine intragastric ad- ministration 1 time per day at a dose of 50 mg per 100 g body weight for two weeks. There was also assessed the dynamics of tumor growth amongst the animals (III or control group, n = 8) receiving siRNA without chan- ging the functional state of thyroid gland. To obtain the tumor suspension we preliminary se- lected some cells of the tumor by its incubation with col- lagenase (with concentration 1 µl/ml) for 1 h at 37 oC. Afterwards, an additional splitting of the tumor cells was done with the automated system for mechanical tissue disaggregation «BD Medimachine» (Germany). The tu- mor cells were filtered with a syringe through the cap Steri-Dual (3 M «Health Care» (Germany)). Next, the cells were counted in the Goryaev’s chamber. Then RPMI 1640 medium was added up to the concentration of (5–6) �106 cells/ml. 0.5 ml of the received tumor sus- pension was administered subcutaneously to the rats of the Group IA and IIB. The transfection mixture was prepared separately. 100 µl DMEM medium with high glucose concentra- tion was added to a sterile tube with 4 µl of transfection reagent Turbofect R-0537 («Fermentas», Lithuania) and then the diluted transfection reagent was kept at room temperature for 10–20 min. The diluted transfection re- agent was added with 1 µl siRNA («Dharmacon», USA, P-011511-07-0005 siRNA, human SOCS1). Then they were gently stirred with a pipette, and incubated at room temperature for 10–20 min to form transfection comple- xes. The obtained mixture was diluted with 500 µl of cul- ture medium RPMI 1640. Then the culture medium was aspirated from the tumor cells and immediately repla- ced with a mixture of the diluted reagent of transfection. In 20–30 min the rats of Group IIA and IIB were inoculated with the tumor cells with siRNA. Within 21 days the development of the tumors in the experimental rats was observed. Then animals were decapitated under a mild chloroform anesthesia. The specimens were stained with hematoxylin and eosin and analyzed with optical microscope Leica DM750 (Ger- many) with photo-video output, camera (5 M pixels). We carried out immunohistochemical studies of proliferation marker Ki-67. Proliferation index was cal- culated as the proportion of positively stained nuclei of tumor cells within the proliferative compartment in 5 randomly selected fields of view (> 500 cells) [7, 8]. Statistical analysis was performed using the soft- ware Statistica 6.15 («StatSoft Inc.», USA) [9]. Results and discussion. The siRNA transfection which is the specific inhibition of endogenous genes sig- nificantly affected the tumor growth dynamics (Table). It could be concluded that siRNA transfection redu- ced tumor growth in hypothyroid rats by 25.5 %, while the tumor growth in hyperthyroid status decreased only by 3.5 %. This makes possible to assume the thyroid hormone ability to block siRNA inhibitory effect. During the histological examination of tumors deri- ved from experimental animals with simulated hypothy- roid state it was found that the tumor areas consisted of polymorphic cells with the signs of glandular epitheli- um, large hyperchromatic nucleus, coarse chromatin, 464 ZAPOROZHAN V. M. ET AL. homogenous eosinophilic cytoplasm and clear outlines. A shape of nuclei was uneven with indrawings. Tumor cells were located separately, their nuclei were displa- ced to the periphery. Large areas of destroyed cells with nuclei lysis were observed among the tumor tissue, as well as the fragments of dead cells, dramatically dystro- phic granulocytes. Necrosis occupied approximately 1/3 of the investigated tumor area. Eosinophils dominated over granulocytes. Outgrowth contacts with the forma- tion of lace like structures between the tumor cells were found in the areas with a more compact arrangement of tumor cells. These cells had an amphiphilic cytoplasm, their borders were uneven. A large number of capillaries with flattened endothelium were observed among the tu- mor tissue. Necrosis of tumor cells was observed in non- vascular and pericapillary space. After the siRNA transfection in tumor tissue of hy- pothyroid rats, destroyed cells with nuclei lysis, cell- shading, fibrin strands and mixed dystrophic granulo- cytes were frequently found among the fragments of dead cells. Necrosis of tumor cells also was observed in non-vascular and pericapillary space (Fig. 1, see inset). On the other hand, somewhat different picture was observed in the hyperthyroid state. In a histological sec- tion of the encapsulated tumor site which consisted of large polymorphic cells with signs of glandular epithe- lium and large hyperchromatic nucleus there were diffe- rent variants of the nuclei location: from central to fully displaced to the periphery. The nuclei had a coarse chro- matin with the presence of 1–3 large nucleoli. The sha- pe of most nuclei was smooth, concave-oval. The cells contained homogeneous amphiphilic cytoplasm with clear boundaries. The tumor cells were placed separate- ly at greater area, they had a nucleus shifted to the peri- phery. There were observed the outgrowth contacts with the formation of lace like and perivasculary palisade li- ke structures between the tumor cells. Among the tumor tissue there were also found small line areas of destroy- ed tumor cells with nuclei lysis, the fragments of necro- tic cells mixed with sharply dystrophic granulocytes and the fragments of necrotic granulocytes. The nec- rosis fields occupied about 1/5 of all the investigated tumor area. At the level of tumor cells with necrosis the- re were a small number of isolated convex-oval tumor cells with the eccentric rounded nucleus and eosinophi- lic cytoplasm. There were large number of capillaries with flattened endothelium. After application of siRNA transfection of animal’s tumor at hyperthyroidism we observed slightly diffe- rent histological figure (Fig. 2, see inset). There were do- minated the areas with relatively compact arrangement of tumor cells with outgrowth contacts and the forma- tion of lace like structures. The cytoplasm of tumor cells was amphiphilic. The boundaries of cells were uneven and with outgrowing. Parts of the nuclei had a rough form with indrawings. Other areas consisted of separa- tely located polymorphic cells with a large hyperchro- mic nucleus. Nucleus was shifted to the periphery. It was defined a coarse chromatin and a homogeneous eosino- philic cytoplasm with clear bounds. There were also ob- served the fields of destroyed cells with nuclei lysis, fal- ling out of fibrin strands and admixture of dramatically dystrophic granulocytes among the fragments of dead cells. Besides, there were the following changes: the compact areas of tumor cells changed to more isolated and then to necrosis. A large number of capillaries were 465 IMPACT OF siRNA TRANSFECTION ON TUMOR DEVELOPMENT Group Days of the experiment 9 12 15 18 21 ²À 2.38 ± 0.15 7.76 ± 0.37* 15.82 ± 0.55* 18.77 ± 0.61* 21.39 ± 0.41* ²B 2.6 ± 0.13* 6.22 ± 0.25* 12.92 ± 0.45* 17.35 ± 0.39* 19.22 ± 0.74* ²²À 1.71 ± 0.08 5.5 ± 0.15 8.02 ± 0.25* 11.41 ± 0.44 15.92 ± 0.48 ²²B 1.62 ± 0.14 5.18 ± 0.17 9.5 ± 0.31 14.83 ± 0.64 18.53 ± 0.72 ²II (control) 1.78 ± 0.09 5.18 ± 0.17 10.72 ± 0.41 13.5 ± 0.46 16.52 ± 0.48 Note. *The difference with the control group is statistically significant (P < 0.05). Dynamics of tumor growth (cm3) in the experimental groups (M ± m) ISSN 0233-7657. Biopolymers and Cell. 2013. Vol. 29. N 6 Figures to article by V. M. Zaporozhan et al. a b Fig. 1. Histology of the modified pathomorphosis in a hypothyroid state: a – without the siRNA transfection, � 400; b – with the siRNA transfection, � 200. Hematoxylin-eosin a b Fig. 2. Histology of the modified pathomorphosis in a hyperthyroid state: a – without the siRNA transfection, �400; b – with siRNA transfection, � 100. Hematoxylin-eosin among the tumor, some of them had blood stasis and mi- xed thrombi without evidence of organization. The to- tal volume of necrosis was about 1/4 of the total tissue, necrosis fields were larger in the depth of tumor site and lesser – under the capsule of connective tissue. The bor- der of the tumor with the granulation tissue was clear. The index of tumor cells proliferation in female- rats with simulated hypothyroidism in the group IA was 14.4 %, whereas in the group IIA it was only 5.6 %. The proliferation index has not changed after siRNA transfection introduction in the model of experimental hyperthyroidism: in the IB group it was 10.3 %, and in IIB – 9.6 % (p > 0.05). While studying oncogenesis much attention has be- en recently paid to the signal transducer and activator of transcription 3 (STAT3). STAT3 is classified as a proto-oncogene because its activated form can mediate oncogenic transformation in cells culture and induce the formation of tumors [10]. Several studies have shown that STAT3 is activated in many cancers, such as leuke- mia, gliomas, cancer of the head and neck, melanoma, breast cancer, prostate cancer, cervical cancer, endomet- rial cancer [11]. Tyrosine kinases Jak2, onkoprotein Src and vascular endothelial growth factor (VEGF) are potential activators of STAT3. Abnormally activated STAT3 activates a number of genes which manage the behavior of tumor cells (survival ability, growth, angio- genesis, invasion and resistance to immune oversight). The suppressor of cytokine signaling (SOCS1) is one of the genes which can reduce the tyrosine kinase activity and block Jak2 leading to the inhibition of STAT3 phosphorylation [12]. Activation of SOCS1 is influenced by cytokines. This gene inhibits hormonal signals and suppresses cytokine signals acting via the intracellular negative feedback reducing the activity of tyrosine kinase Jak connected with the receptor [12]. There is information that siRNA also can inhibit proliferation and increase apoptosis as a result of inter- ruption of STAT3 phosphorylation through reducing regulatory Jak2 activation [13]. siRNA also suppresses the expression of VEGF, which regulates angiogenesis, and the tumor immune response in cancer. As a result of this interaction, the tumor cell proliferation decreases, apoptosis and expression of both mature and immature markers of dendritic cells (HLA-DR, CD80, CD86, CD40, CD1a, CD83) increase [14]. Thus, the results obtained strongly prove that the tu- mor inhibition effect of small interfering RNA takes place to a greater extent at hypothyroid state than at hy- perthyroidism. This phenomenon indicates an important role of thyroid hormones in the regulation of the genes expression, which controls the cell cycle. According to the literature data [15], we can make an assumption that thyroxine activates the proliferation of tumor cells mo- re than their apoptosis. The prevalence of necrobiotic processes and characteristic changes in the blood vessels may be associated with the specific effects of thyroid hormones on angiogenesis. In addition to the gene expression inhibition, one more interesting property of siRNA is the ability to trig- ger independently an immune response – to activate al- pha- and beta-interferons, cytokines and other media- tors of inflammation [16]. So, besides the ability to suppress the expression of genes which trigger tumor development, siRNA can al- so directly activate the expression of SOCS1 gene and induce antitumor immune response. This property pro- vides siRNA a great potential to be used in genetic the- rapy of cancer. Conclusions. After inoculation of Guerin’s carci- noma to female-rats with simulated hypo- and hyper- thyroidal states the proliferation of tumor cells is more active in the group of animals with experimental hypo- thyroidism. The siRNA trasfection leads to the inhibition of tu- mor growth in animals with both hypo- and hyperthy- roidism, but this process is more active at hypothy- roidism. The prospects for further research are associated with assessment of a possibility to use siRNA transfec- tion for control and activation of apoptosis in an expe- rimental model of tumorogenesis. Â. Ì. Çàïîðîæàí, Ã. Ñ. Ìàðèíþê, O. Ë. Õîëîäêîâà, Â. Â. Áóáíîâ, Ä. Þ. Àíäðîíîâ Îö³íêà ìîæëèâîñò³ ñóïðåñ³¿ ïóõëèííîãî ðîñòó ³ç âèêîðèñòàííÿì òðàíñôåêö³¿ ³íòåðôåðóþ÷èõ ÐÍÊ SOCS1 íà ïðèêëàä³ åêñïåðèìåíòàëüíî¿ ìîäåë³ óòâîðåííÿ ïóõëèíè çà çì³íåíî¿ ôóíêö³¿ ùèòîïîä³áíî¿ çàëîçè Ðåçþìå Ìåòà. Îö³íèòè âïëèâ òðàíñôåêö³éíî ââåäåíî¿ siÐÍÊ íà ðîçâèòîê ïóõëèíè çà óìîâ åêñïåðèìåíòàëüíî¿ ïàòîëîã³¿ ùèòîïîä³áíî¿ çàëî- çè. Ìåòîäè. Åêñïåðèìåíòè âèêîíóâàëè íà ñàìèöÿõ ùóð³â ìàñîþ 466 ZAPOROZHAN V. M. ET AL. 180 ± 20 ã. Òâàðèí ðîçä³ëåíî íà ï’ÿòü ãðóï: IA ³ IÁ – ùóðè ç ìîäåëü- îâàíèìè ã³ïî- ³ ã³ïåðòèðåîçîì òà ïåðåâèòîþ êàðöèíîìîþ Ãåðå- íà, IIA ³ IIÁ – ç ìîäåëüîâàíèìè ã³ïî- ³ ã³ïåðòèðåîçîì òà ïåðåâè- òîþ êàðöèíîìîþ Ãåðåíà ó êîìïëåêñ³ ç òðàíñôåêö³éíî ââåäåíîþ siÐÍÊ, ²²² – ç ïåðåâèòîþ êàðöèíîìîþ Ãåðåíà ó êîìïëåêñ³ ç òðàíñ- ôåêö³éíî ââåäåíîþ siÐÍÊ. Âèì³ðþâàëè îðòîãîíàëüí³ ðîçì³ðè ïóõ- ëèíè. Ïðîâîäèëè ã³ñòîëîã³÷í³ òà ³ìóíîã³ñòîõ³ì³÷í³ äîñë³äæåííÿ çðàçê³â ïóõëèíè. Ðåçóëüòàòè. Ïîêàçàíî, ùî ³íã³áóâàëüíèé âïëèâ êîðîòêèõ ³íòåðôåðóþ÷èõ ÐÍÊ á³ëüøîþ ì³ðîþ ïðîÿâëÿºòüñÿ çà ã³ïîòèðåî¿äíîãî ñòàíó, ùî ñâ³ä÷èòü ïðî âàæëèâó ðîëü ãîðìîí³â ùèòîïîä³áíî¿ çàëîçè ó ðåãóëÿö³¿ åêñïðåñ³¿ ãåí³â, êîíòðîëþþ÷èõ êë³òèííèé öèêë. Îáãîâîðþºòüñÿ éìîâ³ðí³ñòü çàëåæíîñò³ ïåðåâà- æàííÿ íåêðîá³îòè÷íèõ ïðîöåñ³â òà õàðàêòåðíèõ çì³í ó ñóäèíàõ â³ä ä³¿ òèðåî¿äíèõ ãîðìîí³â íà àíã³îãåíåç ³ ïðîë³ôåðàòèâí³ ïðîöå- ñè. Âèñíîâêè. Òðàñôåêö³ÿ siÐÍÊ ïðèçâîäèòü äî ïðèãí³÷åííÿ ïóõ- ëèííîãî ðîñòó ó òâàðèí ÿê ç ã³ïî-, òàê ³ ç ã³ïåðòèðåîçîì, ïðîòå àêòèâí³øå öåé ïðîöåñ ïðîò³êàº ïðè ã³ïîòèðåî¿äíîìó ñòàí³. Êëþ÷îâ³ ñëîâà: òðàíñôåêö³ÿ, s³ÐÍÊ, åêñïåðèìåíòàëüíà ìî- äåëü, ã³ïîòèðåîç, ã³ïåðòèðåîç. Â. Ì. Çàïîðîæàí, À. Ñ. Ìàðèíþê, Å. Ë. Õîëîäêîâà, Â. Â. Áóáíîâ, Ä. Þ. Àíäðîíîâ Îöåíêà âîçìîæíîñòè ñóïðåññèè îïóõîëåâîãî ðîñòà ñ èñïîëüçîâàíèåì òðàíñôåêöèè èíòåðôåðèðóþùèõ ÐÍÊ SOCS1 íà ïðèìåðå ýêñïåðèìåíòàëüíîé ìîäåëè îáðàçîâàíèÿ îïóõîëè ïðè èçìåíåííîé ôóíêöèè ùèòîâèäíîé æåëåçû Ðåçþìå Öåëü. Îöåíêà âëèÿíèÿ òðàíñôåêöèîííî ââåäåííîé siÐÍÊ íà ðàç- âèòèå îïóõîëè â óñëîâèÿõ ýêñïåðèìåíòàëüíîé ïàòîëîãèè ùèòî- âèäíîé æåëåçû. Ìåòîäû. Ýêñïåðèìåíòû ïðîâîäèëè íà ñàìêàõ êðûñ ìàññîé 180 ± 20 ã. Æèâîòíûõ ðàçäåëèëè íà ïÿòü ãðóïï: IA è IÁ – êðûñû ñ ìîäåëèðîâàííûìè ãèïî- è ãèïåðòèðåîçîì è ïåðåâè- òîé êàðöèíîìîé Ãåðåíà, IIA è IIÁ – ñ ìîäåëèðîâàííûìè ãèïî- è ãè- ïåðòèðåîçîì è ïåðåâèòîé êàðöèíîìîé Ãåðåíà â êîìïëåêñå ñ òðàíñôåêöèîííî ââåäåííîé siÐÍÊ, III – êîíòðîëüíàÿ ãðóïïà ñ ïå- ðåâèòîé êàðöèíîìîé Ãåðåíà â êîìïëåêñå ñ òðàíñôåêöèîííî ââå- äåííîé siÐÍÊ. Èçìåðÿëè îðòîãîíàëüíûå ðàçìåðû îïóõîëè. Îñóùå- ñòâëÿëè ãèñòîëîãè÷åñêèå è èììóíîãèñòîõèìè÷åñêèå èññëåäîâà- íèÿ îáðàçöîâ îïóõîëè. Ðåçóëüòàòû. Ïîêàçàíî, ÷òî èíãèáèðóþ- ùåå âëèÿíèå êîðîòêèõ èíòåðôåðèðóþùèõ ÐÍÊ â áîëüøåé ñòå- ïåíè ïðîÿâëÿåòñÿ ïðè ãèïîòèðåîèäíîì ñîñòîÿíèÿ, ÷òî óêàçûâà- åò íà âàæíóþ ðîëü ãîðìîíîâ ùèòîâèäíîé æåëåçû â ðåãóëÿöèè ýêñ- ïðåññèè ãåíîâ, êîíòðîëèðóþùèõ êëåòî÷íûé öèêë. Îáñóæäàåòñÿ âåðîÿòíîñòü çàâèñèìîñòè ïðåîáëàäàíèÿ íåêðîáèîòè÷åñêèõ ïðî- öåññîâ è õàðàêòåðíûõ èçìåíåíèé â ñîñóäàõ îò âëèÿíèÿ òèðåîèä- íûõ ãîðìîíîâ íà àíãèîãåíåç è ïðîëèôåðàòèâíûå ïðîöåññû. Âû- âîäû. Òðàíñôåêöèÿ siÐÍÊ ïðèâîäèò ê óãíåòåíèþ îïóõîëåâîãî ðîñòà ó æèâîòíûõ êàê ñ ãèïî-, òàê è ñ ãèïåðòèðåîçîì, îäíàêî áî- ëåå àêòèâíî ýòîò ïðîöåññ ïðîòåêàåò ïðè ãèïîòèðåîèäíîãî ñî- ñòîÿíèè. Êëþ÷åâûå ñëîâà: òðàíñôåêöèÿ, s³ÐÍÊ, ýêñïåðèìåíòàëüíàÿ ìîäåëü, ãèïîòèðåîç, ãèïåðòèðåîç. REFERENCES 1. Brent G. A. Mechanisms of thyroid hormone action // J. Clin. In- vest.–2012.–122, N 9.–Ð. 3035–3043. 2. Sar P., Peter R., Rath B. Das Mohapatra A., Mishra S. K. 3, 3'5 Triiodo L thyronine induces apoptosis in human breast cancer MCF-7 cells, repressing SMP30 expression through negative thyroid response elements // PLoS One.–2011.–6, N 6.–e20861. 3. Khatri N., Rathi M., Baradia D., Trehan S., Misra A. In vivo de- livery aspects of miRNA, shRNA and siRNA // Crit. Rev. Ther. Drug. Carrier. 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Impact of siRNA transfection on tumor development under experimental pathology of thyroid gland

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