Apoptosis indices in liver cell mitochondria in B16/F10 melanoma growing in presence of chronic neurogenic pain

2021 ◽  
pp. 71-79
Author(s):  
Е.М. Франциянц ◽  
И.В. Нескубина ◽  
Е.И. Сурикова ◽  
А.И. Шихлярова ◽  
В.А. Бандовкина ◽  
...  
Keyword(s):  
Cytochrome C ◽  
G Protein ◽  
Tumor Development ◽  
Mitochondrial Fraction ◽  
Liver Mitochondria ◽  
Caspase 9 ◽  
Neurogenic Pain ◽  
Intact Group ◽  
Subcutaneous Transplantation ◽  
Melanoma Growth

Введение. Печень по количеству, плотности митохондрий один из самых богатых органов, который также является критическим местом для множества метаболических путей. Цель исследования - изучение показателей апоптоза в митохондриях печени самок мышей линии С57ВL/6 при самостоятельном росте меланомы В16/F10 и на фоне коморбидной патологии - хронической нейрогенной боли. Методика. В эксперименте использовали мышей-самок (n=168) линии С57ВL/6. Группы: интактная (n=21); контрольная (n=21) - создание модели хронической нейрогенной боли (ХНБ), путем двусторонней перевязки седалищных нервов; группа сравнения (n=63) - подкожная трансплантация меланомы B16/F10; основная группа (ХНБ+B16/F10) (n=63) - подкожная трансплантация меланомы В16/F10 через 3 нед после моделировия ХНБ. В митохондриях печени методом ИФА определяли концентрацию: цитохрома С (нг/г белка), каспазы 9 (нг/г белка), Bcl-2 (нг/г белка), AIF (нг/г белка), кальция (Са 2+) (мМоль/г белка). Результаты. В митохондриях клеток печени через 1 нед роста меланомы относительно интактных значений фиксировали нарастание уровней AIF в 2,2 раза, цитохрома С в 1,7 раза (р<0,05) и снижение каспазы 9 в 2,0 раза; через 3 нед - падение кальция в 4,7 раза, AIF в 7,1 раза и цитохрома С в 1,7 раза (р<0,05) и накопление каспазы 9 - 1,6 раза (р<0,05). Развитие опухоли при ХНБ через 1 нед сопровождалось уменьшением концентрации AIF в 29,3 раза и цитохрома С в 2,0 раза по сравнению с контрольными значениями (ХНБ). Через 3 недели роста меланомы на фоне ХНБ фиксировали снижение уровней AIF в 6,6 раза, цитохрома С в 4,7 раза и кальция в 32,8 раза, уровень каспазы 9, напротив, повышался в 1,5 раза (р<0,05). Заключение. Наличие коморбидной патологии - ХНБ при опухолевом процессе способствует раннему возникновению нарушений в электронно-транспортной цепи митохондрий клеток печени. Background. The liver is one of the richest organs in terms of the number and density of mitochondria; it is also a critical site for many metabolic pathways. The aim of the study was to analyze indicators of apoptosis in liver mitochondria in female С57ВL/6 mice with B16/F10 melanoma growing alone and in presence of chronic neurogenic pain. Methods. Female С57ВL/6 mice (n=168) were studied. Animals were divided into groups: intact group (n=21); controls (n=21) with a model of chronic neurogenic pain (CNP) created by bilateral sciatic nerve ligation; comparison group (n=63) with subcutaneous transplantation of B16/F10 melanoma; main group (CNP+B16/F10) (n=63) with subcutaneous transplantation of B16/F10 melanoma 3 wks after modeling CNP. Cytochrome C (ng/g protein), caspase-9 (ng/g protein), Bcl-2 (ng/g protein), AIF (ng/g protein), and calcium (Ca2+) (mmol/g protein) were measured by ELISA in the liver mitochondrial fraction. Results. After 1 wk of melanoma growth, AIF increased by 2.2 times, cytochrome C increased by 1.7 times (p<0.05), and caspase-9 decreased by 2.0 times compared to the intact group values. After 3 wks, calcium decreased by 4.7 times, AIF by 7.1 times, cytochrome C by 1.7 times (p<0.05), and caspase-9 increased by 1.6 times (p<0.05). After 1 wk, tumor development in the presence of CNP was accompanied by decreases in AIF by 29.3 times and cytochrome C by 2.0 times, compared to control CNP values. After 3 wks of melanoma growth in presence of CNP, AIF decreased by 6.6 times, cytochrome C by 4.7 times, and calcium by 32.8 times. Caspase-9, on the contrary, increased by 1.5 times (p<0.05). Conclusions. The presence of CNP comorbidity during the tumor development facilitates earlier occurrence of disorders in the electron transport chain of hepatocyte mitochondria.

scholarly journals The state of apoptosis factor system in mitochondria of skin and tumor cells in standard and stimulated growth of B16/F10 melanoma in female C57BL/6 mice

2021 ◽  
Vol 8 (1) ◽  
pp. 8-19
Author(s):  
E. M. Frantsiyants ◽  
I. V. Neskubina ◽  
E. I. Surikova ◽  
A. I. Shikhlyarova ◽  
I. V. Kaplieva ◽  
...  
Keyword(s):  
Cytochrome C ◽  
Tumor Growth ◽  
Tumor Cells ◽  
Caspase 9 ◽  
Neurogenic Pain ◽  
Skin Cells ◽  
Similar Tendency ◽  
Thermo Fisher Scientific ◽  
Fisher Scientific ◽  
Melanoma Growth

Purpose of the study. Studying the dynamics of factors of apoptosis in mitochondria of skin and tumors cells in female mice with melanoma growth stimulated by chronic neurogenic pain. Material and methods. The study included female С57ВL/6 mice (n=56) with a model of chronic neurogenic pain (CNP) produced by the bilateral sciatic nerve ligation and with transplanted B16/F10 melanoma. After 1–3 weeks of the tumor growth, levels of cytochrome C, caspase‑9 (Bioscience, Austria), Bcl‑2 (Thermo Fisher Scientific, Austria), and AIF (RayBiotech, USA) were determined by ELISA, and levels of calcium (Са2+) were determined by the Arsenazo III method (Abris+, Russia) in mitochondria of tumors cells and skin not affected by the tumor growth. Results. In the CNP state, mitochondria of the skin cells showed a significant increase in Са2+ by 96.7 times, AIF by 1.4 times and Bcl‑2 by 5.9 times, while caspase‑9 decreased by 2.6 times, compared to the levels in intact mice. In the CNP‑stimulated melanoma growth, mitochondria of cells of the skin not affected by the tumor growth demonstrated a decrease in all studied indices, except caspase‑9 – its levels increased by 4.6 times after 3 weeks of the tumor growth. In mitochondria of the tumor cells within 1–3 weeks, levels of Са2+ decreased over time by 37.2–96.1 times, respectively, AIF by 49.4–2.0 times, Bcl‑2 by 3.0–1.5 times, cytochrome C by 15.3–8.8 times, and caspase‑9 increased by 1.7–4.4 times compared with the level in animals with pain. Conclusions. In general, the growth of melanoma stimulated by chronic pain and the standard melanoma growth were characterized by the opposite dynamics of levels of apoptosis factor both in mitochondria of skin cells and in mitochondria of tumor cells, with the exception of cytochrome C. Mitochondria of melanoma cells and of the unchanged skin have a similar tendency to change the levels of apoptosis factors, which may indicate their functioning in the conditions of the mitochondrial network at the level of one organ. Mitochondria of tumor cells provide the anti‑apoptotic state of the tumor itself and of the skin not affected by the malignant process, probably due to the stress state of the skin.

scholarly journals Influence of B16/F10 melanoma growth variant on calcium levels in mitochondria in various organs of female mice

2021 ◽  
Vol 8 (1) ◽  
pp. 20-29
Author(s):  
O. I. Kit ◽  
E. M. Frantsiyants ◽  
I. V. Neskubina ◽  
E. I. Surikova ◽  
I. V. Kaplieva ◽  
...  
Keyword(s):  
Tumor Growth ◽  
Tumor Cells ◽  
Pain Syndrome ◽  
Neurogenic Pain ◽  
Intact Group ◽  
Female Mice ◽  
Terminal Stage ◽  
Initial Stage ◽  
The Brain ◽  
Melanoma Growth

Purpose of the study. To analyze the calcium levels in mitochondria of cells in different organs in standard and stimulated growth of experimental В16/F10 melanoma. Materials and Methods. The study included female С57ВL/6 mice (n=168). Experimental groups: intact group (n=21), group with a model of chronic neurogenic pain (CNP) (n=21), group M – B16/F10 melanoma (n=63), group M+CNP – mice (n=63) with transplantation of B16/F10 melanoma 3 weeks after CNP model creation. The concentration of calcium in mitochondrial samples was determined by a biochemical method (Abris+, Russia). Results were statistically analyzed using the Statistica 10.0 program. Results. CNP decreased calcium levels in mitochondria of cells in the brain by 1.4 (р=0.00153) times, liver by 2.6 times and heart by 3.2 times and increased the levels in the skin by 97.1 times. In standard growth of experimental melanoma, levels of calcium in cell mitochondria in most of the studied organs increased at the initial stage of the melanoma growth, and decreased to intact values and lower by the terminal stage. In the mitochondria of tumor cells, calcium levels were stably high at all stages of standard tumor growth. At the initial stage of CNP‑stimulated tumor growth, a decrease in calcium in the mitochondria of the skin by 5.7 times and its accumulation in the mitochondria of the brain by 6.6 times, heart, and kidneys were recorded by 1.5 times. At the terminal stage of stimulated melanoma growth, extremely low calcium values were recorded in the mitochondria of all organs. A stably low level of calcium was registered in the mitochondria of tumor cells at all stages of stimulated melanoma growth. Conclusions. The growth of experimental B16/F10 melanoma in female mice is accompanied by mitochondrial dysfunction affecting most organs. Stimulation of the growth of experimental melanoma with chronic neurogenic pain, unlike the standard growth variant, changes accumulation of calcium in the mitochondria of cells both in organs and in the tumor itself. The chronic pain syndrome accompanying a malignant process can influence its course with the involvement of mitochondria and the modification of their functions.

scholarly journals Influence of a B16/F10 melanoma variant on the Вcl-2 levels in mitochondria in various organs of female mice

2021 ◽  
Vol 20 (3) ◽  
pp. 46-53
Author(s):  
O. I. Kit ◽  
E. M. Frantsiyants ◽  
I. V. Neskubina ◽  
N. D. Cheryarina ◽  
A. I. Shikhlyarova ◽  
...  
Keyword(s):  
Tumor Growth ◽  
Tumor Development ◽  
Growth Period ◽  
Pathological Process ◽  
Intact Group ◽  
Kidney Mitochondria ◽  
Female Mice ◽  
Thermo Fisher Scientific ◽  
Liver And Kidney ◽  
Melanoma Growth

Aim. To study the Bcl-2 level in mitochondria of various organs in female mice with standard and stimulated growth of an experimental B16/F10 melanoma.Materials and methods. The study included С57ВL/6 female mice (n = 168). The experimental animals were divided into the following groups: an intact group (n = 21), a group with modelled chronic neuropathic pain (CNP) (n = 21), an M group with B16/F10 melanoma (n = 63), and a CNP + M group (n = 63). The Bcl-2 concentration (ng / mg protein) in mitochondrial samples was determined by ELISA (Thermo Fisher Scientific, Austria). Statistical analysis of the results was carried out using Statistica 10.0.Results. Compared to the Bcl-2 levels in the intact animals, CNP decreased this parameter in the cardiac mitochondria by 1.3 times, while increasing it by 5.9 times in the skin mitochondria. In the dynamics of standard melanoma growth, the Bcl-2 content changed compared with the corresponding intact values in the mitochondria of the brain, heart, and skin, but did not change in the liver and kidneys. In the mitochondria in melanoma, the Bcl-2 levels were high throughout the entire period of standard tumor growth in comparison with the intact skin. The stimulated melanoma growth in CNP was involving more organs into the pathological process as the tumor was growing. Thus, in comparison with the values in the CNP group, the mitochondrial Bcl-2 levels changed in the heart at week 1; in the heart and skin – at week 2; in the heart, skin, and brain – at week 3. The Bcl-2 levels did not change in the liver and kidney mitochondria. In the mitochondria in the CNP-stimulated melanoma, the Bcl-2 levels were lower than in the skin mitochondria in CNP throughout the entire tumor growth period.Conclusion. The liver and kidney mitochondria are somewhat Bcl-2 stable in both standard and stimulated tumor growth. It is assumed that different Bcl-2 dynamics in the mitochondria in melanoma depending on the variant of tumor development reflects the modulating effect of CNP and the ability to change the Bcl-2 levels according to the growth phase.

Comorbidity affects Вcl-2 levels in mitochondria in C57BL/6 mice with transplantable B16/F10 melanoma.

2021 ◽  
Vol 39 (15_suppl) ◽  
pp. e21581-e21581
Author(s):  
Irina V. Neskubina ◽  
Elena M. Frantsiyants ◽  
Valeria A. Bandovkina ◽  
Natalia D. Cheryarina ◽  
Alla I. Shikhlyarova ◽  
...  
Keyword(s):  
Tumor Growth ◽  
Tumor Development ◽  
Brain Mitochondria ◽  
The Body ◽  
Heart Mitochondria ◽  
Control Group ◽  
Neurogenic Pain ◽  
Intact Group ◽  
Vital Functions ◽  
Thermo Fisher Scientific

e21581 Background: Overexpression of the Bcl-2 protein inhibits apoptosis and promotes carcinogenesis. Stress causes signaling leading to cell buffering with Bcl-2 protein above acceptable levels. The purpose of the study was to analyze the influence of comorbidity – chronic neurogenic pain (CNP) – on the Bcl-2 levels in mitochondria of cells of melanoma, the heart, skin and brain in female mice with growing tumors. Methods: Female С57ВL/6 mice were divided into groups: intact group (n = 21); control group with a CNP model – bilateral sciatic nerve ligation (n = 21); group M – B16/F10 melanoma (n = 63); CNP+M group – B16/F10 melanoma was transplanted 3 weeks after the CNP model creation (n = 63). The concentration of Bcl-2 (ng/mg of protein) was determined in mitochondrial samples by ELISA (Thermo Fisher Scientific, Austria). Statictical analysis of results: Statistica 10.0. Results: CNP decreased the Bcl-2 level in heart mitochondria by 1.3 times (p < 0.05), but increased it in skin and brain mitochondria by 5.8 and 1.3 times, respectively. Similar changes were observed in melanoma growth 1 week after its transplantation: Bcl-2 levels decreased in heart mitochondria by 1.3 times, and increased in the skin and brain by 8.9 and 1.3 times, respectively. After 2 weeks of the tumor growth, Bcl-2 in brain mitochondria decreased by 1.7 times, and it started declining in the skin by the 3rd week – by 4 times, compared to intact females. Bcl-2 in tumor mitochondria exceeded the values in the skin by more than 4 times throughout the experiment. Tumor growth in presence of CNP caused a decrease in Bcl-2 in brain mitochondria by 2.4 times after 3 weeks, and in the heart and skin – by 2 and 1.7 times, respectively, after 2 weeks. Bcl-2 in tumor mitochondria in presence of CNP was lower than in the intact skin on average by 1.8 times throughout the experiment. Conclusions: CNP as a comorbidity caused a modulating effect on the mechanisms of survival and apoptosis of cells both in the tumor and in the main organs providing the vital functions of the body - the brain and heart, and also affects the target organ of melanoma - the skin. The results demonstrated the ability of comorbidity to change levels of Bcl-2 in mitochondria depending on the stage of tumor development.

scholarly journals Functional state of cardiomyocyte mitochondria in malignant process in presence of comorbid pathology in experiment

2021 ◽  
Vol 2 (3) ◽  
pp. 13-22
Author(s):  
E. M. Frantsiyants ◽  
I. V. Neskubina ◽  
N. D. Cheryarina ◽  
E. I. Surikova ◽  
A. I. Shikhlyarova ◽  
...  
Keyword(s):  
Diabetes Mellitus ◽  
Cytochrome C ◽  
Free Radical Oxidation ◽  
Control Group ◽  
Heart Cells ◽  
Neurogenic Pain ◽  
Intact Group ◽  
Guerin’S Carcinoma ◽  
Group 2 ◽  
Group 1

Purpose of the study. An analysis of indices of free radical oxidation and respiration of mitochondria of heart cells in a malignant process in presence of diabetes mellitus and chronic neurogenic pain in experimental animals.Materials and methods. The study included outbred female rats (n=32) and С57ВL/6 female mice (n=84). Experimental groups of rats were: intact group 1 (n=8), control group 1 (n=8) with diabetes mellitus (DM), comparison group 1 (n=8) with standard subcutaneous transplantation of Guerin’s carcinoma, main group 1 (n=8) with Guerin’s carcinoma transplanted after 1 week of persistent hyperglycemia. Experimental groups of mice were: intact group 2 (n=21), control group 2 (n=21) with a model of chronic neurogenic pain (CNP), comparison group 2 (n=21) with standard subcutaneous transplantation of melanoma (B16/F10), main group 2 (n=21) (CNP+B16/F10) with melanoma transplanted 3 weeks after the CNP model creation. Heart mitochondria were isolated by differential centrifugation. Levels of cytochrome C (ng/mg of protein), 8-hydroxy-2'-deoxyguanosine (8-OHdG) (ng/mg of protein), and malondialdehyde (MDA) (μmol/g of protein) were measured in mitochondrial samples by ELISA. Statistical analysis was performed using the Statistica 10.0 program.Results. DM in rats upregulated 8-OHdG by 6.3 times and MDA by 1.9 times (р=0.0000) and downregulated cytochrome C by 1.5 times (р=0.0053) in heart cell mitochondria, compared to intact values. DM+Guerin’s carcinoma in rats increased 8-OHdG by 14.0 times and MDA by 1.7 times (р=0.0000) and decreased cytochrome C by 1.5 times (р=0.0000), compared to intact values. CNP in mice did not affect the studied parameters in mitochondria of the heart. CNP+B16/F10 in mice increased 8-OHdG by 7.1 times and MDA by 1.6 times (р=0.0000) and decreased cytochrome C by 1.6 times (р=0.0008).Conclusions. Comorbidity (diabetes mellitus, chronic neurogenic pain) together with malignant pathology aggravates mitochondrial dysfunction of heart cells with destabilization of the respiratory chain mediated by free radical oxidation processes.

scholarly journals Influence of B16/F10 melanoma growth variant on the level of cytochrome C in mitochondria in various organs of female mice

2021 ◽  
Vol 27 (4) ◽  
pp. 46-52
Author(s):  
E. M. Frantsiyants ◽  
I. V. Neskubina ◽  
N. D. Cheryarina ◽  
E. I. Surikova ◽  
L. A. Nemashkalova
Keyword(s):  
Statistical Analysis ◽  
Cytochrome C ◽  
Complex Iii ◽  
Neurogenic Pain ◽  
Complex Iv ◽  
Female Mice ◽  
Slowing Down ◽  
Analysis Of Results ◽  
The Brain ◽  
Melanoma Growth

Introduction. Cytochrome C in mitochondria transfers electrons from complex III to complex IV, and it is a signaling molecule in the apoptosis realization.The objective was to evaluate the level of cytochrome C in cell mitochondria in various organs of female mice with standard and stimulated growth of experimental B16/F10 melanoma.Methods and materials. The experiment was performed on female C57BL/6 mice (n=168). The groups were: intact animals (n=21); controls with a model of chronic neurogenic pain (CNP) (n = 21); group M — standard B16/F10 melanoma transplantation (n=63), group CNP + M — B16/F10 melanoma transplantation 3 weeks after CNP model creation (n=63). The level of cytochrome C (ng / mg protein) were measured by ELISA (Bioscience, Austria). Statistical analysis of results was performed using the «Statistica 10.0» program.Results. After 1 week of standard melanoma growth, an increase in the level of cytochrome C by 2.7 and 1.7 times was detected in mitochondria of the brain and liver; by the 3rd week, it decreased in the liver and skin by 1.7 times. In melanoma mitochondria, the level of cytochrome C was lower than in the skin of intact animals: by 2.5 times after week 1, by 4.5 times after week 2, and by 4.6 times after week 3. After 1 week of stimulated melanoma growth, the level of cytochrome C decreased compared control values: by 2.2 times in the brain, by 1.9 times in the liver, by 1.4 times in the skin; by week 3, it decreased by 4.8 times in mitochondria of the brain, by 4.7 times — in the liver, by 2.3 times — in the heart, by 1.9 times — in the skin. In melanoma mitochondria, the level of cytochrome C was lower than in the skin of intact animals: by 15.3 times after week 1, by 10.3 times after week 2, and by 8.8 times after week 3.Conclusion. Low level of cytochrome C were found in melanoma mitochondria in standard and stimulated tumor growth. The data can be used in the experiment and in clinic for using exogenous cytochrome C as an agent slowing down the malignant process.

Sign in / Sign up

Export Citation Format

Share Document