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Published in: Pituitary 3/2020

01-06-2020

Effects of octreotide on autophagy markers and cell viability markers related to metabolic activity in rat pituitary tumor cells

Authors: Giovanni Tulipano, Andrea Giustina

Published in: Pituitary | Issue 3/2020

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Abstract

Purpose

The aim of this work was to investigate possible direct effects of the somatostatin analog octreotide on autophagy markers and markers of cellular metabolic activity using in vitro cultured rat pituitary tumor cells (GH3 cell line).

Methods

We measured two markers of the autophagic flux in cell lysates by Western blot and MTT reductive activity, total cellular ATP levels, pyruvate dehydrogenase (PDH) complex activity in cells lysates as markers of cell viability related to metabolic activity.

Results

Octreotide (100 nM) treatment induced autophagy activation (increased LC3-I protein lipidation) and enhanced the autophagic flux (SQSTM1/p62 protein downregulation) in GH3 cells in different incubation media, in detail in Hank’s balanced salt solution (HBSS) as well as in maintenance medium with serum. We did not observe any decrease of redox activity and energy production related to the induction of autophagy by octreotide. On the other hand, short-term treatments with octreotide in HBSS tended to enhance MTT reduction activity and to increase PDH complex enzymatic activity and ATP levels measured in GH3 cell lysates.

Conclusions

We provided evidence that octreotide can affect autophagy in pituitary tumor cells. The observed effects of octreotide were not related to a decrease of cellular metabolic activity. Finally, the induction of autophagy was either short-lived or overshadowed by other factors in the long term and this limit does not help clarifying their real impact on the pharmacological activity of somatostatin analogs.
Literature
1.
go back to reference Shintani T, Klionsky DJ (2004) Autophagy in health and disease: a double-edged sword. Science 306:990–995CrossRef Shintani T, Klionsky DJ (2004) Autophagy in health and disease: a double-edged sword. Science 306:990–995CrossRef
2.
go back to reference Mizushima N (2007) Autophagy: process and function. Genes Dev 21:2861–2873CrossRef Mizushima N (2007) Autophagy: process and function. Genes Dev 21:2861–2873CrossRef
3.
go back to reference Mizushima N, Levine B, Cuervo AM, Klionsky DJ (2008) Autophagy fights disease through cellular self-digestion. Nature 7182:1069–1075CrossRef Mizushima N, Levine B, Cuervo AM, Klionsky DJ (2008) Autophagy fights disease through cellular self-digestion. Nature 7182:1069–1075CrossRef
4.
go back to reference Mizushima N, Yoshimori T, Levine B (2010) Methods in mammalian autophagy research. Cell 140:313–324CrossRef Mizushima N, Yoshimori T, Levine B (2010) Methods in mammalian autophagy research. Cell 140:313–324CrossRef
5.
go back to reference Weckman A, Rotondo F, Di Ieva A, Syro LV, Butz H, Cusimano MD, Kovacs K (2015) Autophagy in endocrine tumors. Endocr Relat Cancer 22:R205–R218CrossRef Weckman A, Rotondo F, Di Ieva A, Syro LV, Butz H, Cusimano MD, Kovacs K (2015) Autophagy in endocrine tumors. Endocr Relat Cancer 22:R205–R218CrossRef
6.
go back to reference Morselli E, Galluzzi L, Kepp O, Vicencio JM, Criollo A, Maiuri MC, Kroemer G (2009) Anti- and pro-tumor functions of autophagy. Biochem Biophys Acta 1793:1524–1532CrossRef Morselli E, Galluzzi L, Kepp O, Vicencio JM, Criollo A, Maiuri MC, Kroemer G (2009) Anti- and pro-tumor functions of autophagy. Biochem Biophys Acta 1793:1524–1532CrossRef
7.
go back to reference Weckman A, Di Ieva A, Rotondo F, Syro LV, Ortiz LD, Kovacs K, Cusimano MD (2014) Autophagy in the endocrine glands. J Mol Endocrinol 52:151–163CrossRef Weckman A, Di Ieva A, Rotondo F, Syro LV, Ortiz LD, Kovacs K, Cusimano MD (2014) Autophagy in the endocrine glands. J Mol Endocrinol 52:151–163CrossRef
8.
go back to reference Mercau ME, Repetto EM, Perez MN, Martinez Calejman C, Sanchez Puch S, Finkielstein CV, Cymeryng CB (2016) Moderate exercise prevents functional remodeling of the anterior pituitary gland in diet-induced insulin resistance in rats: role of oxidative stress and autophagy. Endocrinology 157:1135–1145CrossRef Mercau ME, Repetto EM, Perez MN, Martinez Calejman C, Sanchez Puch S, Finkielstein CV, Cymeryng CB (2016) Moderate exercise prevents functional remodeling of the anterior pituitary gland in diet-induced insulin resistance in rats: role of oxidative stress and autophagy. Endocrinology 157:1135–1145CrossRef
9.
go back to reference Dagistanli FK, Ozkaya HM, Kucukyoruk B, Biceroglu H, Metin D, Gazioglu N, Oz B, Kadioglu P, Ozturk M (2018) Preoperative somatostatin analogue treatment might trigger apoptosis and autophagy in tumor tissues of patients with acromegaly: a pilot study. Exp Clin Endocrinol Diabetes 126:168–175CrossRef Dagistanli FK, Ozkaya HM, Kucukyoruk B, Biceroglu H, Metin D, Gazioglu N, Oz B, Kadioglu P, Ozturk M (2018) Preoperative somatostatin analogue treatment might trigger apoptosis and autophagy in tumor tissues of patients with acromegaly: a pilot study. Exp Clin Endocrinol Diabetes 126:168–175CrossRef
10.
go back to reference Kun Z, Yuling Y, Dongchun W, Bingbing X, Xiaoli L, Bin X (2016) HIF-I alpha inhibition sensitized pituitary adenoma cells to temozolomide by regulating presenilin I expression and autophagy. Technol Cancer Res Treat 15:NP95–NP104. Kun Z, Yuling Y, Dongchun W, Bingbing X, Xiaoli L, Bin X (2016) HIF-I alpha inhibition sensitized pituitary adenoma cells to temozolomide by regulating presenilin I expression and autophagy. Technol Cancer Res Treat 15:NP95–NP104.
11.
go back to reference Lin SJ, Leng ZG, Guo YH, Cai L, Cai Y, Li N, Shang HB, Le WD, Zhao WG, Wu ZB (2015) Suppression of mTOR pathway and induction of autophagy-dependent cell death by cabergoline. Oncotarget 17:39329–39341 Lin SJ, Leng ZG, Guo YH, Cai L, Cai Y, Li N, Shang HB, Le WD, Zhao WG, Wu ZB (2015) Suppression of mTOR pathway and induction of autophagy-dependent cell death by cabergoline. Oncotarget 17:39329–39341
12.
go back to reference Melmed S, Bronstein MD, Chanson P, Klibanski A, Casanueva FF, Wass JAH, Strasburger CJ, Luger A, Clemmons DR, Giustina A (2018) A consensus Statement on acromegaly therapeutic outcomes. Nat Rev 14:552–561 Melmed S, Bronstein MD, Chanson P, Klibanski A, Casanueva FF, Wass JAH, Strasburger CJ, Luger A, Clemmons DR, Giustina A (2018) A consensus Statement on acromegaly therapeutic outcomes. Nat Rev 14:552–561
13.
go back to reference Amato R, Catalani E, Dal Monte M, Cammalleri M, Di Renzo I, Perrotta C, Cervia D, Casini G (2018) Autophagy-mediated neuroprotection induced by octreotide in an ex vivo model of early diabetic retinopathy. Pharmacol Res 128:167–178CrossRef Amato R, Catalani E, Dal Monte M, Cammalleri M, Di Renzo I, Perrotta C, Cervia D, Casini G (2018) Autophagy-mediated neuroprotection induced by octreotide in an ex vivo model of early diabetic retinopathy. Pharmacol Res 128:167–178CrossRef
14.
go back to reference Sun H, Zou S, Candiotti KA, Peng Y, Zhang Q, Xiao W, Wen Y, Wu J, Yang J (2017) Octreotide attenuates acute kidney injury after hepatic ischemia and reperfusion by enhancing autophagy. Sci Rep 7:42701CrossRef Sun H, Zou S, Candiotti KA, Peng Y, Zhang Q, Xiao W, Wen Y, Wu J, Yang J (2017) Octreotide attenuates acute kidney injury after hepatic ischemia and reperfusion by enhancing autophagy. Sci Rep 7:42701CrossRef
15.
go back to reference Takahashi S, Abe T, Gotoh J, Fukuuchi Y (2002) Substrate-dependence of reduction of MTT: a tetrazolium dye differs in cultured astroglia and neurons. Neurochem Int 40:441–448CrossRef Takahashi S, Abe T, Gotoh J, Fukuuchi Y (2002) Substrate-dependence of reduction of MTT: a tetrazolium dye differs in cultured astroglia and neurons. Neurochem Int 40:441–448CrossRef
16.
go back to reference Günther T, Tulipano G, Dournaud P, Bousquet C, Csaba Z, Kreienkamp HJ, Lupp A, Korbonits M, Castaño JP, Wester HJ, Culler M, Melmed S, Schulz S (2018) International union of basic and clinical pharmacology CV Somatostatin receptors: structure, function, ligands, and new nomenclature. Pharmacol Rev 70:763–835. Günther T, Tulipano G, Dournaud P, Bousquet C, Csaba Z, Kreienkamp HJ, Lupp A, Korbonits M, Castaño JP, Wester HJ, Culler M, Melmed S, Schulz S (2018) International union of basic and clinical pharmacology CV Somatostatin receptors: structure, function, ligands, and new nomenclature. Pharmacol Rev 70:763–835.
17.
go back to reference Tulipano G, Faggi L, Schulz S, Spinello M, Giustina A (2017) Effects of pasireotide (SOM230) on protein turnover and p70S6 kinase-S6 ribosomal protein signaling pathway in rat skeletal muscle cells. Endocrine 57:179–182CrossRef Tulipano G, Faggi L, Schulz S, Spinello M, Giustina A (2017) Effects of pasireotide (SOM230) on protein turnover and p70S6 kinase-S6 ribosomal protein signaling pathway in rat skeletal muscle cells. Endocrine 57:179–182CrossRef
18.
go back to reference Peverelli E, Lania AG, Mantovani G, Beck-Peccoz P, Spada A (2009) Characterization of intracellular signalling mediated by human somatostatin receptor 5: role of the DRY motif and the third intracellular loop. Endocrinology 150:3169–3176CrossRef Peverelli E, Lania AG, Mantovani G, Beck-Peccoz P, Spada A (2009) Characterization of intracellular signalling mediated by human somatostatin receptor 5: role of the DRY motif and the third intracellular loop. Endocrinology 150:3169–3176CrossRef
19.
go back to reference Theodoropoulou M, Zhang J, Laupheimer S, Paez-Pereda M, Erneux C, Florio T, Pagotto U, Stalla GK (2006) Octreotide, a somatostatin analogue, mediates its antiproliferative action in pituitary tumor cells by altering phosphatidylinositol 3-kinase signaling and inducing zac1 expression. Can Res 66:1576–1582CrossRef Theodoropoulou M, Zhang J, Laupheimer S, Paez-Pereda M, Erneux C, Florio T, Pagotto U, Stalla GK (2006) Octreotide, a somatostatin analogue, mediates its antiproliferative action in pituitary tumor cells by altering phosphatidylinositol 3-kinase signaling and inducing zac1 expression. Can Res 66:1576–1582CrossRef
20.
go back to reference Cheung NW, Boyages SC (1995) Somatostatin-14 and its analog octreotide exert a cytostatic effect on GH3 rat pituitary tumor cell proliferation via a transient G0/G1 cell cycle block. Endocrinology 136:4174–4181CrossRef Cheung NW, Boyages SC (1995) Somatostatin-14 and its analog octreotide exert a cytostatic effect on GH3 rat pituitary tumor cell proliferation via a transient G0/G1 cell cycle block. Endocrinology 136:4174–4181CrossRef
21.
go back to reference Tulipano G, Faggi L, Losa M, Mortini P, Spinello M, Sibilia V, Pagani F, Cocchi D, Giustina A (2013) Effects of AMPK activation and combined treatment with AMPK activators and somatostatin on hormone secretion and cell growth in cultured GH-secreting pituitary tumor cells. Mol Cell Endocrinol 365:197–206CrossRef Tulipano G, Faggi L, Losa M, Mortini P, Spinello M, Sibilia V, Pagani F, Cocchi D, Giustina A (2013) Effects of AMPK activation and combined treatment with AMPK activators and somatostatin on hormone secretion and cell growth in cultured GH-secreting pituitary tumor cells. Mol Cell Endocrinol 365:197–206CrossRef
22.
go back to reference Klionsky DJ et al (2012) Guidelines for the use and interpretation of assays for monitoring autophagy. Autophagy 8:445–544CrossRef Klionsky DJ et al (2012) Guidelines for the use and interpretation of assays for monitoring autophagy. Autophagy 8:445–544CrossRef
23.
go back to reference Carroll B, Dunlop EA (2017) The lysosome: a crucial hub for AMPK and mTORC1 signalling. Biochem J 474:1453–1466CrossRef Carroll B, Dunlop EA (2017) The lysosome: a crucial hub for AMPK and mTORC1 signalling. Biochem J 474:1453–1466CrossRef
24.
go back to reference Rardin MJ, Wiley SE, Naviaux RK, Murphy AN, Dixon JE (2009) Monitoring phosphorylation of the pyruvate dehydrogenase complex. Anal Biochem 389:157–164CrossRef Rardin MJ, Wiley SE, Naviaux RK, Murphy AN, Dixon JE (2009) Monitoring phosphorylation of the pyruvate dehydrogenase complex. Anal Biochem 389:157–164CrossRef
25.
go back to reference Linher-Melville K, Singh G (2017) The complex roles of STAT3 and STAT5 in maintaining redox balance: Lessons from STAT-mediated xCT expression in cancer cells. Mol Cell Endocrinol 451:40–52CrossRef Linher-Melville K, Singh G (2017) The complex roles of STAT3 and STAT5 in maintaining redox balance: Lessons from STAT-mediated xCT expression in cancer cells. Mol Cell Endocrinol 451:40–52CrossRef
26.
go back to reference Yao-Xia L, Jing-Yan C, Xia-Lian T, Ping C, Min Z (2017) The 20 kDa and 22 kDa forms of human growth hormone (hGH) exhibit different intracellular signalling profiles and properties. Gen Comp Endocrinol 248:49–54CrossRef Yao-Xia L, Jing-Yan C, Xia-Lian T, Ping C, Min Z (2017) The 20 kDa and 22 kDa forms of human growth hormone (hGH) exhibit different intracellular signalling profiles and properties. Gen Comp Endocrinol 248:49–54CrossRef
27.
go back to reference Liu G, Robillard L, Banihashemi B, Albert PR (2002) Growth hormone-induced diacylglycerol and ceramide formation via GαI3 and Gβγ in GH4 pituitary cells. Potentiation by dopamine-D2 receptor activation. J Biol Chem 277:48427–48433CrossRef Liu G, Robillard L, Banihashemi B, Albert PR (2002) Growth hormone-induced diacylglycerol and ceramide formation via GαI3 and Gβγ in GH4 pituitary cells. Potentiation by dopamine-D2 receptor activation. J Biol Chem 277:48427–48433CrossRef
28.
go back to reference Jeoung NH (2015) Pyruvate Dehydrogenase Kinases: therapeutic targets for diabetes and cancers. Diabetes Metab J 39:188–197CrossRef Jeoung NH (2015) Pyruvate Dehydrogenase Kinases: therapeutic targets for diabetes and cancers. Diabetes Metab J 39:188–197CrossRef
29.
go back to reference Giustina A, Mazziotti G, Torri V, Spinello M, Floriani I, Melmed S (2012) Meta-analysis on the effects of octreotide on tumor mass in acromegaly. PLoS ONE 7(5):e36411CrossRef Giustina A, Mazziotti G, Torri V, Spinello M, Floriani I, Melmed S (2012) Meta-analysis on the effects of octreotide on tumor mass in acromegaly. PLoS ONE 7(5):e36411CrossRef
30.
go back to reference Mazziotti G, Giustina A (2010) Effects of lanreotide SR and Autogel on tumor mass in patients with acromegaly: a systematic review. Pituitary 13:60–67CrossRef Mazziotti G, Giustina A (2010) Effects of lanreotide SR and Autogel on tumor mass in patients with acromegaly: a systematic review. Pituitary 13:60–67CrossRef
31.
go back to reference Colao A, Grasso LFS, Giustina A, Melmed S, Chanson P, Pereira AM, Pivonello R (2019) Acromegaly. Nat Rev 5(1):20 Colao A, Grasso LFS, Giustina A, Melmed S, Chanson P, Pereira AM, Pivonello R (2019) Acromegaly. Nat Rev 5(1):20
32.
go back to reference Gola M, Bonadonna S, Mazziotti G, Amato G, Giustina A (2006) Resistance to somatostatin analogs in acromegaly: an evolving concept? J Endocrinol Investig 29:86–93CrossRef Gola M, Bonadonna S, Mazziotti G, Amato G, Giustina A (2006) Resistance to somatostatin analogs in acromegaly: an evolving concept? J Endocrinol Investig 29:86–93CrossRef
33.
go back to reference Giustina A, Bronstein MD, Chanson P, Petersenn S, Casanueva FF, Sert C, Houchard A, Melmed S (2019) Staging and managing patients with acromegaly in clinical practice: baseline data from the SAGIT® validation study. Pituitary 22:476–487CrossRef Giustina A, Bronstein MD, Chanson P, Petersenn S, Casanueva FF, Sert C, Houchard A, Melmed S (2019) Staging and managing patients with acromegaly in clinical practice: baseline data from the SAGIT® validation study. Pituitary 22:476–487CrossRef
Metadata
Title
Effects of octreotide on autophagy markers and cell viability markers related to metabolic activity in rat pituitary tumor cells
Authors
Giovanni Tulipano
Andrea Giustina
Publication date
01-06-2020
Publisher
Springer US
Published in
Pituitary / Issue 3/2020
Print ISSN: 1386-341X
Electronic ISSN: 1573-7403
DOI
https://doi.org/10.1007/s11102-020-01028-0

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