Modulation of proteomic profile in H295R adrenocortical cell line induced by mitotane

Stigliano, Antonio; Cerquetti, Lidia; Borro, Marina; Gentile, Giovanna; Bucci, Barbara; Misiti, Silvia; Piergrossi, P.; Brunetti, Ercole; Simmaco, Maurizio; Toscano, Vincenzo

doi:10.1677/erc-07-0003

Cited by 41 publications

(34 citation statements)

References 31 publications

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“…As already noted, mitotane, at concentrations lower than those reached in vivo, is able to reduce steroids and TSH secretion in adrenocortical and pituitary TSHsecreting cell lines respectively (Stigliano et al 2008. Our findings support the evidence that mitotane affects pituitary function, also reducing the secretory activity of corticotroph cells.…”

Section: Discussionsupporting

confidence: 91%

Mitotane reduces human and mouse ACTH-secreting pituitary cell viability and function

Gentilin¹,

Tagliati²,

Terzolo³

et al. 2013

Journal of Endocrinology

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Medical therapy for Cushing's disease (CD) is currently based on agents mainly targeting adrenocortical function. Lately, pituitary-directed drugs have been developed, with limited efficacy. Mitotane, a potent adrenolytic drug, has been recently investigated for the treatment of CD, but the direct pituitary effects have not been clarified so far. The aim of our study was to investigate whether mitotane may affect corticotroph function and cell survival in the mouse pituitary cell line AtT20/D16v-F2 and in the primary cultures of human ACTHsecreting pituitary adenomas, as an in vitro model of pituitary corticotrophs. We found that in the AtT20/D16v-F2 cell line and in primary cultures, mitotane reduces cell viability by inducing caspase-mediated apoptosis and reduces ACTH secretion. In the AtT20/D16v-F2 cell line, mitotane reduces Pomc expression and blocks the stimulatory effects of corticotropinreleasing hormone on cell viability, ACTH secretion, and Pomc expression. These effects were apparent at mitotane doses greater than those usually necessary for reducing cortisol secretion in Cushing's syndrome, but still in the therapeutic window for adrenocortical carcinoma treatment. In conclusion, our results demonstrate that mitotane affects cell viability and function of human and mouse ACTH-secreting pituitary adenoma cells. These data indicate that mitotane could have direct pituitary effects on corticotroph cells.

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Section: Discussionsupporting

confidence: 91%

Mitotane reduces human and mouse ACTH-secreting pituitary cell viability and function

Gentilin¹,

Tagliati²,

Terzolo³

et al. 2013

Journal of Endocrinology

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“…Nevertheless, these effects were observed only at doses as high as 168 mM, whereas 80 mM MTT was not effective on either mitochondrial structure or steroid secretion (Fang 1979). It may be hypothesized that low intracellular levels of MTT or short treatment durations affect primarily mitochondrial steroidogenic enzymatic activity with little effects on cellular viability (Stigliano et al 2008, Lin et al 2012. Conversely, higher doses of MTT or longer treatment durations cause a mitochondrial toxic effect, in addition to reduced steroid hormone secretion (Fang 1979, present paper), resulting in cell death (Cai et al 1995b, present paper).…”

Section: Discussionmentioning

confidence: 90%

“…In particular, mitochondrial enzymes, but not microsomal ones, in the steroidogenic pathways (StAR, CYP11A1, and CYP11B1) have been demonstrated to be inhibited in the H295 and H295R cells (Stigliano et al 2008, Asp et al 2010, Lin et al 2012. Different strains of H295 cells have shown different degrees of inhibition in steroid secretion, depending on their steroidogenic profiles (Samandari et al 2007).…”

Section: Discussionmentioning

confidence: 99%

“…In the treated cells, we observed a net reduction of oxygen consumption in the mitochondria-rich fraction (nearly 80% inhibition with 50 mM MTT), which seems to be mainly due to a damage of the mitochondrial outer membrane (a significant decrease in the expression of VDAC is evident) rather than to a specific derangement of the respiratory chain complexes (stable levels of SDH). MTT has been previously demonstrated to downregulate the expression of proteins involved in cell redox potential, cholesterol trafficking, and stress response as well as in the maintenance of mitochondrial integrity (Stigliano et al 2008). Reduction in the expression of VDAC1 following MTT treatment may not be only a passive consequence of the disruption of the mitochondrial ultrastructure and increased permeability of the outer membrane.…”

Section: Discussionmentioning

confidence: 99%

“…Some early sporadic studies have reported the qualitative effects of MTT on animal adrenals (Kaminsky et al 1962, Moore et al 1980, Krüger et al 1984 or on the human atypical estrogen-secreting adrenal cancer cell line Fang-8 (Fang 1979). Recently, by evaluating MTT-induced variation of the proteomic profile of the ACC H295R cell line (Stigliano et al 2008), MTT has been suggested to interact with some proteins belonging to the classes of energetic metabolism, stress response, cytoskeletal structure, and tumorigenesis. Thus, the aim of the present study was to evaluate the potential toxic activity of MTT and its cellular targets in human cell models of ACC.…”

Section: Introductionmentioning

confidence: 99%

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Morphofunctional effects of mitotane on mitochondria in human adrenocortical cancer cells

Poli¹,

Guasti²,

Rapizzi³

et al. 2013

Endocrine-Related Cancer

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At present, mitotane (MTT) represents the first-line pharmacological approach for the treatment of advanced adrenocortical carcinoma (ACC). Despite clear evidence that the drug can reduce the clinical signs of steroid excess in secreting ACC, the mechanism mediating the possible toxic effect of MTT on tumor cells still remains obscure. This study investigated the intracellular events underlying the toxic effect of MTT by studying qualitative and quantitative alterations in mitochondrial morphology and functions in human adrenocortical cancer cell lines, H295R and SW13. Increasing concentrations of MTT resulted in rapid intracellular accumulation and conversion of the drug. Cytostatic and cytotoxic effects were evident at doses corresponding to the therapeutic window (30-50 mM) through an apoptotic mechanism involving caspase 3/7. Electron microscopic analysis of cell mitochondria displayed MTT-induced dose-and time-dependent alterations in the morphology of the organelle. These alterations were characterized by a marked swelling and a decrease in the number of respiratory cristae, accompanied by a significant depolarization of the mitochondrial membrane potential, finally leading to the disruption of the organelle. A drastic reduction of oxygen consumption was observed due to mitochondrial membrane damage, which was accompanied by a decrease in the levels of VDAC1 integral membrane channel. These findings contribute to better understand the intracellular mechanism of action of MTT in ACC cells, showing that its cytotoxic effect seems to be mainly mediated by an apoptotic process activated by the disruption of mitochondria.

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MAPK/ERK pathway inhibition is a promising treatment target for adrenocortical tumors

Pereira

Monteiro

Costa

et al. 2018

J of Cellular Biochemistry

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Unraveling molecular mechanisms that regulate tumor development and proliferation is of the utmost importance in the quest to decrease the high mortality rate of adrenocortical carcinomas (ACC). Our aim was to evaluate the role of two of the mitogen-activated protein kinase (MAPK) signaling pathways (extracellular signal-regulated protein kinases [ERKs 1/2] and p38) in the adrenocortical tumorigenesis, as well as the therapeutic potential of MAPK/ERK inhibition. ERKs 1/2 and p38 activation were evaluated in incidentalomas (INC; n = 10), benign Cushing's syndrome (BCS; n = 12), malignant Cushing's syndrome (MCS; n = 6) and normal adrenal glands (NAG; 8). ACC cell line (H295R) was used to evaluate the ability of PD184352 (0.1, 1, and 10 µM), a specific MEK-MAPK-ERK pathway inhibitor, to modulate cell proliferation, viability, metabolism, and steroidogenesis. ERKs 1/2 activation was significantly higher in MCS (2.83 ± 0.17) compared with NAG (1.00 ± 0.19 "arbitrary units"), INC (1.20 ± 0.13) and BCS (2.09 ± 0.09). Phospho-p38 expression was absent in all the MCS analyzed. MAPK/ERK kinase (MEK) inhibition with PD184352 significantly decreased proliferation as well as steroidogenesis and also increased the redox state of the H295R cells. This data suggests that MEK-MAPK-ERK signaling has a role in adrenocortical tumorigenesis that could be potentially used as a diagnostic marker for malignancy and targeted treatment in ACC.

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Modulation of proteomic profile in H295R adrenocortical cell line induced by mitotane

Cited by 41 publications

References 31 publications

Mitotane reduces human and mouse ACTH-secreting pituitary cell viability and function

Mitotane reduces human and mouse ACTH-secreting pituitary cell viability and function

Morphofunctional effects of mitotane on mitochondria in human adrenocortical cancer cells

MAPK/ERK pathway inhibition is a promising treatment target for adrenocortical tumors

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