cAMP-dependent protein kinase activation inhibits proliferation and enhances apoptotic effect of tumor necrosis factor-α in NCI-H295R adrenocortical cells

Liu, Jiangi; Li, X. D.; Ora, Ari; Heikkilä, Päivi; Vaheri, Antti; Voutilainen, Raimo

doi:10.1677/jme.1.01535

Cited by 19 publications

(10 citation statements)

References 45 publications

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“…upregulated or downregulated c-Myc level did not affect significantly cell growth in the cell lines. our data are in agreement with Liu and colleagues (44) who showed that the overexpression of C-MYC by transient transfection assay was not correlated with proliferation in H295R cell line. This finding shows how other factors involved in the C-MYC pathway are essential for the occurrence to proliferative effect in cancer.…”

Section: Discussionsupporting

confidence: 93%

C-MYC modulation induces responsiveness to paclitaxel in adrenocortical cancer cell lines

Cerquetti

Sampaoli

Salvo

et al. 2015

International Journal of Oncology

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Abstract. C-MYC is overexpressed in many types of cancer linked to poor prognosis. We examined the c-Myc protein expression in adrenocortical cancer (ACC) cells to investigate the role of this protein in the neoplasm, its involvement in chemotherapy and finally to determine whether c-Myc could be considered a prognostic factor in patients with ACC. H295R and SW13 cell lines were treated with paclitaxel. c-Myc overexpressing cell clones were achieved by transfecting the H295R cell line with the pcDNA3-hMYC plasmid expressing the full-lengh C-MYC coding sequence. The SW13 cell line was transfected with siRNA oligonucleotides for C-MYC. Cell cycle analysis was evaluated by flow cytometry. c-Myc, cyclin B1 and pro caspase expression levels were evaluated by western blot analysis. We found that expression of c-Myc was highly expressed in the SW13 cells, whereas the protein was undetectable in the H295R cells. Different doses of paclitaxel were required in the two ACC cell line to induce a block in the G 2 phase, characterized by increased cyclin B1 levels and to induce apoptosis by procaspase-3 activation. Interestingly, the silencing of C-MYC mRNA prevented paclitaxel induced apoptosis in SW13 cells, whereas in the H295R cells the overexpression of C-MYC rendered the cells more prone to growth inhibition after paclitaxel exposure. The present study directly demonstrates that C-MYC plays a central role in controlling proliferation in ACC cells after paclitaxel treatment and that c-Myc could be considered as a marker for predicting response to chemotherapeutic agents in ACC cell lines. IntroductionAdrenocortical carcinoma (ACC) is a rare endocrine neoplasia with a variable prognosis, depending on tumor stage and time of diagnosis, but it is generally fatal, with an overall survival of 5 years from detection (1-3). Metastasis associated with ACCs can range from 30 to 85% (4).More recently, elevated or deregulated expression of the c-Myc protein has been detected in a wide range of human cancers, and is often associated with aggressive and poorly differentiated tumors. C-MYC belongs to the class of immediate early genes which are induced in response to a number of mitogenic signals. It encodes a nuclear transcription factor leading to activation or repression of target genes, thus affecting several biological processes, such as cellular growth, differentiation, cell cycle and apoptosis (5,6). Surprisingly, gene expression profile studies have demonstrated an underexpression of C-MYC in ACC compared to adrenocortical adenoma (7-10).The treatment with DNA-damaging agents can sensitize cells to apoptosis when c-Myc is overexpressed (11), but its role in cellular susceptibility to anticancer drugs is controversial. It has been reported that the overexpression of c-Myc not only enhances tumor cell sensitivity (12) but also induces resistance to antineoplastic agents (13).It has been widely demonstrated that cells expressing high c-Myc levels show an apoptosis-prone phenotype in response to different stimuli, affecting cell c...

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Section: Discussionsupporting

confidence: 93%

C-MYC modulation induces responsiveness to paclitaxel in adrenocortical cancer cell lines

Cerquetti

Sampaoli

Salvo

et al. 2015

International Journal of Oncology

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“…Two cAMP analogs that activate PKA induced differentiation and also reduced proliferation of H295R cells [16]. Altogether these findings support a model of PPNAD in which PRKAR1A mutation decreases PKA-I activity that in turn leads to IGFBP-2 induction and enhanced adrenocortical cell proliferation.…”

Section: Discussionsupporting

confidence: 76%

“…For example, ERK1/2 inhibition by PKA has been implicated in the latter's inhibition of cellular proliferation, and this inhibition was reversed in Carney complex cells [18]. c-myc was induced by PKA activation in adrenocortical cells and decreased their proliferation [16]. Finally, PRKAR1A may have PKA-independent functions [19].…”

Section: Discussionmentioning

confidence: 99%

Primary pigmented nodular adrenocortical disease reveals insulin-like growth factor binding protein-2 regulation by protein kinase A

Shi¹,

Henwood²,

Bannerman³

et al. 2007

Growth Hormone & IGF Research

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Objective-Primary Pigmented Nodular Adrenocortical Disease (PPNAD) can occur as an isolated trait or part of Carney complex, a familial lentiginosis-multiple endocrine neoplasia syndrome frequently caused by mutations in PRKAR1A, which encodes the 1α regulatory subunit of protein kinase A (PKA). Because alterations in the insulin-like growth factor (IGF) axis, particularly IGF-II and IGF binding protein (IGFBP)-2 over-expression, have been implicated in sporadic adrenocortical tumors, we sought to examine the IGF axis in PPNAD.Design-RNA samples and paraffin-embedded sections were procured from adrenalectomy specimens of patients with PPNAD. Changes in expression of IGF axis components were evaluated by real-time quantitative RT-PCR and immunohistochemistry. NCI-H295R cells were used to study PKA and IGF axis signaling in adrenocortical cells in vitro.Results-IGFBP-2 mRNA level distinguished between the two genetic subtypes of this disease; increased IGFBP-2 expression in PRKAR1A mutation-positive PPNAD tissues was also confirmed by immunohistochemistry. Moreover, PKA inhibitors increased IGFBP-2 expression in NCI-H295R adrenocortical cells, and anti-IGFBP-2 antibody reduced their proliferation.Conclusion-IGFBP-2 expression is increased in PPNAD caused by PRKAR1A mutations, and in adrenocortical cancer cells. This is the first evidence for PKA-dependent regulation of IGFBP-2 expression in adrenocortical cells.

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“…The cells were incubated with 1:100-diluted primary antibodies in phosphate-buffered saline (Cav2.2/a1B antibody, Alomone Labs, Jerusalem, Israel; Cav1.2/a1C antibody, Alomone Labs; and Cav3.1/a1G antibody, Sigma-Aldrich) overnight at 4 1C. 27 After being rinsed, the cells were incubated with the secondary antibody (biotinylated anti-rabbit immunoglobulin G; Invitrogen) and the third antibody (streptavidin-conjugated Alexa Fluor 555; Invitrogen) for 1 h at room temperature. Negative controls for each procedure were obtained by replacing the primary antibody with purified nonimmune rabbit immunoglobulin G (1:1000).…”

Section: Immunocytochemical Stainingmentioning

confidence: 99%

Expression of N-type calcium channels in human adrenocortical cells and their contribution to corticosteroid synthesis

et al. 2010

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The inhibition of aldosterone activity is a useful approach for preventing the progression of cardiovascular and renal diseases in hypertensive patients. Although the results of our previous in vivo study suggested that N-type calcium channels may have a role in regulating plasma aldosterone levels, the direct relationship between N-type calcium channels and aldosterone production in adrenocortical cells has not been examined. In this study, the analysis of quantitative reverse transcription-PCR, western blotting, and immunocytological staining indicated the possible presence of N-type calcium channels in human adrenocortical cells (H295R cell line). Patch clamp analysis indicated that omega-conotoxin GVIA (CnTX), an N-type calcium channel inhibitor, suppressed voltage-dependent barium currents. During steroidogenesis, CnTX significantly reduced the transient calcium signaling induced by angiotensin II (Ang II) and partially prevented Ang II-induced aldosterone and cortisol formation with no significant influence on CYP11B2 and CYP11B1 mRNA expression. In addition, in a1B calcium channel subunits, knockdown significantly decreased Ang II-induced aldosterone formation with increments in CYP11B2 mRNA expression. We also investigated the inhibitory activities of some types of dihydropyridine calcium channel blockers (CCBs; cilnidipine: L-/N-type CCB, efonidipine: L-/T-type CCB, and nifedipine: L-type CCB), and these agents showed a dose-dependent inhibition effect on Ang II-induced aldosterone and cortisol production. Furthermore, only cilnidipine failed to suppress CYP11B1 expression in H295R cells. These results suggest that N-type calcium channels have a significant role in transducing the Ang II signal for aldosterone (and cortisol) biosynthesis, which may explain the mechanism by which N-type calcium channels regulate plasma aldosterone levels.

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cAMP-dependent protein kinase activation inhibits proliferation and enhances apoptotic effect of tumor necrosis factor-α in NCI-H295R adrenocortical cells

Cited by 19 publications

References 45 publications

C-MYC modulation induces responsiveness to paclitaxel in adrenocortical cancer cell lines

C-MYC modulation induces responsiveness to paclitaxel in adrenocortical cancer cell lines

Primary pigmented nodular adrenocortical disease reveals insulin-like growth factor binding protein-2 regulation by protein kinase A

Expression of N-type calcium channels in human adrenocortical cells and their contribution to corticosteroid synthesis

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