Plasminogen/plasmin affects expression of glycolysis regulator TIGAR and induces autophagy in lung adenocarcinoma A549 cells

Summary. Pericellular plasmin generation triggers apoptosis/anoikis in normal adherent cells. However, cancer cells are notoriously resistant to anoikis, enabling metastasis and new tumor growth beyond their original environment. Autophagy can be a major contributor to anoikis resistance in cancer....

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Бібліографічні деталі
Дата:2023
Автори: Tykhomyrov, A.A., Nedzvetsky, V.S., Aĝca, C.A., Guzyk, M.M., Korsa, V.V., Grinenko, T.V.
Формат: Стаття
Мова:English
Опубліковано: PH Akademperiodyka 2023
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Онлайн доступ:https://exp-oncology.com.ua/index.php/Exp/article/view/2020-4-4
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Назва журналу:Experimental Oncology

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Experimental Oncology
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institution Experimental Oncology
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datestamp_date 2023-10-11T16:43:48Z
collection OJS
language English
topic anoikis
autophagy
beclin-1
LC3
plasminogen/plasmin
TIGAR
spellingShingle anoikis
autophagy
beclin-1
LC3
plasminogen/plasmin
TIGAR
Tykhomyrov, A.A.
Nedzvetsky, V.S.
Aĝca, C.A.
Guzyk, M.M.
Korsa, V.V.
Grinenko, T.V.
Plasminogen/plasmin affects expression of glycolysis regulator TIGAR and induces autophagy in lung adenocarcinoma A549 cells
topic_facet anoikis
autophagy
beclin-1
LC3
plasminogen/plasmin
TIGAR
anoikis
autophagy
beclin-1
LC3
plasminogen/plasmin
TIGAR
format Article
author Tykhomyrov, A.A.
Nedzvetsky, V.S.
Aĝca, C.A.
Guzyk, M.M.
Korsa, V.V.
Grinenko, T.V.
author_facet Tykhomyrov, A.A.
Nedzvetsky, V.S.
Aĝca, C.A.
Guzyk, M.M.
Korsa, V.V.
Grinenko, T.V.
author_sort Tykhomyrov, A.A.
title Plasminogen/plasmin affects expression of glycolysis regulator TIGAR and induces autophagy in lung adenocarcinoma A549 cells
title_short Plasminogen/plasmin affects expression of glycolysis regulator TIGAR and induces autophagy in lung adenocarcinoma A549 cells
title_full Plasminogen/plasmin affects expression of glycolysis regulator TIGAR and induces autophagy in lung adenocarcinoma A549 cells
title_fullStr Plasminogen/plasmin affects expression of glycolysis regulator TIGAR and induces autophagy in lung adenocarcinoma A549 cells
title_full_unstemmed Plasminogen/plasmin affects expression of glycolysis regulator TIGAR and induces autophagy in lung adenocarcinoma A549 cells
title_sort plasminogen/plasmin affects expression of glycolysis regulator tigar and induces autophagy in lung adenocarcinoma a549 cells
title_alt Plasminogen/plasmin affects expression of glycolysis regulator TIGAR and induces autophagy in lung adenocarcinoma A549 cells
description Summary. Pericellular plasmin generation triggers apoptosis/anoikis in normal adherent cells. However, cancer cells are notoriously resistant to anoikis, enabling metastasis and new tumor growth beyond their original environment. Autophagy can be a major contributor to anoikis resistance in cancer. Aim: To investigate if protective autophagy can be induced in lung adenocarcinoma cells in response to plasminogen treatment. Materials and Methods: Human lung adenocarcinoma A549 cells were incubated with Glu-plasminogen (0.1–1.0 µM) for 24 h. Pericellular plasmin activity was monitored spectrophotometrically by a cleavage of the specific chromogenic­ substrate S-2251. Cell survival was assessed by 3-[4,5-dimethyl thiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT)-test. Degradation of fibronectin, levels of autophagy markers (beclin-1 and light chain 3 (LC3)) and glycolysis regulator (TIGAR) were evaluated by western blot. Intracellular localization of LC-3 was visualized by immunocytochemistry. Results: It was shown that plasminogen is converted into plasmin on the surface of adenocarcinoma cells in a dose-dependent manner. Plasmin disrupted cellular adhesive contacts resulting in cell detachment. A549 cells did not loss their viability after plasminogen treatment for 24 h, while 1.0 µM plasminogen was cytotoxic for non-transformed fibroblasts. Plasminogen 0.1, 0.5, and 1.0 µM induced 7.08-, 5.18-, and 3.78-fold elevation of TIGAR expression (p < 0.05), respectively. Enhanced TIGAR expression indicates switch on pentose phosphate pathway, protection against oxidative stress to prevent apoptosis, facilitation of DNA repair and the degradation of their own organelles (autophagy). Exposure of adenocarcinoma cells to plasminogen in concentrations of 0.1 and 0.5 µM caused 1.74- and 2.19-fold elevation of beclin-1 expression vs untreated cells (p < 0.05), respectively. Unlike K1–3 fragment, plasminogen treatment (0.1-0.5 µM) resulted in increased expression of LC3-I and stimulated rapid conversion of LC3-I to LC3-II. Up-regulation of beclin-1 levels and enhanced LC3-I/II conversion in plasminogen-treated A549 cells are the hallmarks of autophagy induction. According to immunocytochemistry data, increased LC3 puncta and autophagosome formation after exposure to plasminogen could reflect autophagy activation. Conclusions: Therefore, we showed stimulation of prosurvival signals and induction of autophagy in plasminogen-treated adenocarcinoma cells rendering them resistant to apoptosis/anoikis. Based on the obtained data, autophagy has a great potential for novel targets that affect cancer cell death, in addition to the current cytotoxic agents.
publisher PH Akademperiodyka
publishDate 2023
url https://exp-oncology.com.ua/index.php/Exp/article/view/2020-4-4
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spelling oai:ojs2.ex.aqua-time.com.ua:article-1492023-10-11T16:43:48Z Plasminogen/plasmin affects expression of glycolysis regulator TIGAR and induces autophagy in lung adenocarcinoma A549 cells Plasminogen/plasmin affects expression of glycolysis regulator TIGAR and induces autophagy in lung adenocarcinoma A549 cells Tykhomyrov, A.A. Nedzvetsky, V.S. Aĝca, C.A. Guzyk, M.M. Korsa, V.V. Grinenko, T.V. anoikis, autophagy, beclin-1, LC3, plasminogen/plasmin, TIGAR anoikis, autophagy, beclin-1, LC3, plasminogen/plasmin, TIGAR Summary. Pericellular plasmin generation triggers apoptosis/anoikis in normal adherent cells. However, cancer cells are notoriously resistant to anoikis, enabling metastasis and new tumor growth beyond their original environment. Autophagy can be a major contributor to anoikis resistance in cancer. Aim: To investigate if protective autophagy can be induced in lung adenocarcinoma cells in response to plasminogen treatment. Materials and Methods: Human lung adenocarcinoma A549 cells were incubated with Glu-plasminogen (0.1–1.0 µM) for 24 h. Pericellular plasmin activity was monitored spectrophotometrically by a cleavage of the specific chromogenic­ substrate S-2251. Cell survival was assessed by 3-[4,5-dimethyl thiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT)-test. Degradation of fibronectin, levels of autophagy markers (beclin-1 and light chain 3 (LC3)) and glycolysis regulator (TIGAR) were evaluated by western blot. Intracellular localization of LC-3 was visualized by immunocytochemistry. Results: It was shown that plasminogen is converted into plasmin on the surface of adenocarcinoma cells in a dose-dependent manner. Plasmin disrupted cellular adhesive contacts resulting in cell detachment. A549 cells did not loss their viability after plasminogen treatment for 24 h, while 1.0 µM plasminogen was cytotoxic for non-transformed fibroblasts. Plasminogen 0.1, 0.5, and 1.0 µM induced 7.08-, 5.18-, and 3.78-fold elevation of TIGAR expression (p < 0.05), respectively. Enhanced TIGAR expression indicates switch on pentose phosphate pathway, protection against oxidative stress to prevent apoptosis, facilitation of DNA repair and the degradation of their own organelles (autophagy). Exposure of adenocarcinoma cells to plasminogen in concentrations of 0.1 and 0.5 µM caused 1.74- and 2.19-fold elevation of beclin-1 expression vs untreated cells (p < 0.05), respectively. Unlike K1–3 fragment, plasminogen treatment (0.1-0.5 µM) resulted in increased expression of LC3-I and stimulated rapid conversion of LC3-I to LC3-II. Up-regulation of beclin-1 levels and enhanced LC3-I/II conversion in plasminogen-treated A549 cells are the hallmarks of autophagy induction. According to immunocytochemistry data, increased LC3 puncta and autophagosome formation after exposure to plasminogen could reflect autophagy activation. Conclusions: Therefore, we showed stimulation of prosurvival signals and induction of autophagy in plasminogen-treated adenocarcinoma cells rendering them resistant to apoptosis/anoikis. Based on the obtained data, autophagy has a great potential for novel targets that affect cancer cell death, in addition to the current cytotoxic agents. Summary. Pericellular plasmin generation triggers apoptosis/anoikis in normal adherent cells. However, cancer cells are notoriously resistant to anoikis, enabling metastasis and new tumor growth beyond their original environment. Autophagy can be a major contributor to anoikis resistance in cancer. Aim: To investigate if protective autophagy can be induced in lung adenocarcinoma cells in response to plasminogen treatment. Materials and Methods: Human lung adenocarcinoma A549 cells were incubated with Glu-plasminogen (0.1–1.0 µM) for 24 h. Pericellular plasmin activity was monitored spectrophotometrically by a cleavage of the specific chromogenic­ substrate S-2251. Cell survival was assessed by 3-[4,5-dimethyl thiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT)-test. Degradation of fibronectin, levels of autophagy markers (beclin-1 and light chain 3 (LC3)) and glycolysis regulator (TIGAR) were evaluated by western blot. Intracellular localization of LC-3 was visualized by immunocytochemistry. Results: It was shown that plasminogen is converted into plasmin on the surface of adenocarcinoma cells in a dose-dependent manner. Plasmin disrupted cellular adhesive contacts resulting in cell detachment. A549 cells did not loss their viability after plasminogen treatment for 24 h, while 1.0 µM plasminogen was cytotoxic for non-transformed fibroblasts. Plasminogen 0.1, 0.5, and 1.0 µM induced 7.08-, 5.18-, and 3.78-fold elevation of TIGAR expression (p < 0.05), respectively. Enhanced TIGAR expression indicates switch on pentose phosphate pathway, protection against oxidative stress to prevent apoptosis, facilitation of DNA repair and the degradation of their own organelles (autophagy). Exposure of adenocarcinoma cells to plasminogen in concentrations of 0.1 and 0.5 µM caused 1.74- and 2.19-fold elevation of beclin-1 expression vs untreated cells (p < 0.05), respectively. Unlike K1–3 fragment, plasminogen treatment (0.1-0.5 µM) resulted in increased expression of LC3-I and stimulated rapid conversion of LC3-I to LC3-II. Up-regulation of beclin-1 levels and enhanced LC3-I/II conversion in plasminogen-treated A549 cells are the hallmarks of autophagy induction. According to immunocytochemistry data, increased LC3 puncta and autophagosome formation after exposure to plasminogen could reflect autophagy activation. Conclusions: Therefore, we showed stimulation of prosurvival signals and induction of autophagy in plasminogen-treated adenocarcinoma cells rendering them resistant to apoptosis/anoikis. Based on the obtained data, autophagy has a great potential for novel targets that affect cancer cell death, in addition to the current cytotoxic agents. PH Akademperiodyka 2023-05-30 Article Article application/pdf https://exp-oncology.com.ua/index.php/Exp/article/view/2020-4-4 10.32471/exp-oncology.2312-8852.vol-42-no-4.15253 Experimental Oncology; Vol. 42 No. 4 (2020): Experimental Oncology; 270-276 Експериментальна онкологія; Том 42 № 4 (2020): Експериментальна онкологія; 270-276 2312-8852 1812-9269 10.32471/exp-oncology.2312-8852.vol-42-no-4 en https://exp-oncology.com.ua/index.php/Exp/article/view/2020-4-4/2020-4-4 Copyright (c) 2023 Experimental Oncology https://creativecommons.org/licenses/by-nc/4.0/