Onconase Restores Cytotoxicity in Dabrafenib-Resistant A375 Human Melanoma Cells and Affects Cell Migration, Invasion and Colony Formation Capability

Raineri, Alice; Fasoli, Sabrina; Campagnari, Rachele; Gotte, Giovanni; Menegazzi, Marta

doi:10.3390/ijms20235980

Cited by 14 publications

(28 citation statements)

References 54 publications

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“…Nevertheless, the use of a miR-17* mimic and a miR-30c inhibitor significantly decreased the ONC activity and the expression of NFKB1(p50) and its downstream targets [ 78 ]. This result is in agreement with other works that describe a decrease of NFKB1(p50) [ 69 ] and a downregulation in the steady state and subcellular distribution of NF-kappaB [ 66 , 79 , 80 ] as a result of ONC action on the treated cells.…”

Section: Regulatory Rnas Are Key Targets For Different Antitumor Rsupporting

confidence: 93%

Strengths and Challenges of Secretory Ribonucleases as AntiTumor Agents

et al. 2021

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Approaches to develop effective drugs to kill cancer cells are mainly focused either on the improvement of the currently used chemotherapeutics or on the development of targeted therapies aimed at the selective destruction of cancer cells by steering specific molecules and/or enhancing the immune response. The former strategy is limited by its genotoxicity and severe side effects, while the second one is not always effective due to tumor cell heterogeneity and variability of targets in cancer cells. Between these two strategies, several approaches target different types of RNA in tumor cells. RNA degradation alters gene expression at different levels inducing cell death. However, unlike DNA targeting, it is a pleotropic but a non-genotoxic process. Among the ways to destroy RNA, we find the use of ribonucleases with antitumor properties. In the last few years, there has been a significant progress in the understanding of the mechanism by which these enzymes kill cancer cells and in the development of more effective variants. All the approaches seek to maintain the requirements of the ribonucleases to be specifically cytotoxic for tumor cells. These requirements start with the competence of the enzymes to interact with the cell membrane, a process that is critical for their internalization and selectivity for tumor cells and continue with the downstream effects mainly relying on changes in the RNA molecular profile, which are not only due to the ribonucleolytic activity of these enzymes. Although the great improvements achieved in the antitumor activity by designing new ribonuclease variants, some drawbacks still need to be addressed. In the present review, we will focus on the known mechanisms used by ribonucleases to kill cancer cells and on recent strategies to solve the shortcomings that they show as antitumor agents, mainly their pharmacokinetics.

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Section: Regulatory Rnas Are Key Targets For Different Antitumor Rsupporting

confidence: 93%

Strengths and Challenges of Secretory Ribonucleases as AntiTumor Agents

et al. 2021

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“…Preliminary results (not shown) indicated the anti-tumor activity of the dimeric species suffers time latency, similarly to that exerted by ONC in A375 cell line [ 42 , 43 ]. Therefore, we report in Figure 6 A,B the data of cell viability assays measured with sulforhodamine B (SRB) sodium salt after 72 h cells incubation with all RNase A dimers.…”

Section: Resultsmentioning

confidence: 93%

RNase A Domain-Swapped Dimers Produced Through Different Methods: Structure–Catalytic Properties and Antitumor Activity

Montioli

Campagnari

Fasoli

et al. 2021

Life

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Upon oligomerization, RNase A can acquire important properties, such as cytotoxicity against leukemic cells. When lyophilized from 40% acetic acid solutions, the enzyme self-associates through the so-called three-dimensional domain swapping (3D-DS) mechanism involving both N- and/or C-terminals. The same species are formed if the enzyme is subjected to thermal incubation in various solvents, especially in 40% ethanol. We evaluated here if significant structural modifications might occur in RNase A N- or C-swapped dimers and/or in the residual monomer(s), as a function of the oligomerization protocol applied. We detected that the monomer activity vs. ss-RNA was partly affected by both protocols, although the protein does not suffer spectroscopic alterations. Instead, the two N-swapped dimers showed differences in the fluorescence emission spectra but almost identical enzymatic activities, while the C-swapped dimers displayed slightly different activities vs. both ss- or ds-RNA substrates together with not negligible fluorescence emission alterations within each other. Besides these results, we also discuss the reasons justifying the different relative enzymatic activities displayed by the N-dimers and C-dimers. Last, similarly with data previously registered in a mouse model, we found that both dimeric species significantly decrease human melanoma A375 cell viability, while only N-dimers reduce human melanoma MeWo cell growth.

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“…Furthermore, these effects were particularly evident in the A375R cell line, which exhibited resistance to dabrafenib, a mutated BRAF inhibitor, which opens up a new perspective for PARP-i use in the case of resistant melanoma treatment [ 46 ]. Raineri et al showed that A375 cells treated with PARP1-i, AZD2461 in combination with onconase, a ribonuclease enzyme with strong antitumor activity in a number of cancers, inhibited the PARP1-NF-κB pathway, which resulted in reduced cell colony formation, migration, and invasion as well as elevated induction of apoptosis [ 47 , 48 ]. Moreover, another PARP-i, olaparib, although used as a monotherapy, does not exert any significantly therapeutic effect in uveal melanoma patients; however, when combined with dacarbazine, it increased its effectiveness [ 49 ].…”

Section: Discussionmentioning

confidence: 99%

PARP1 as a Marker of an Aggressive Clinical Phenotype in Cutaneous Melanoma—A Clinical and an In Vitro Study

Kupczyk

Simiczyjew

Marczuk

et al. 2021

Cells

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(1) Background: Poly(ADP-ribose) polymerase 1) (PARP1) is a pleiotropic enzyme involved in several cellular processes, e.g., DNA damage repair, regulation of mitosis, and immune response. Little is known about the role of PARP1 in melanoma development and progression. We aimed to investigate the prognostic significance of PARP1 expression in cutaneous melanoma through evaluation of mRNA and protein levels of PARP1 in normal melanocytes and melanoma cell lines, as well as in patients’ tissue material from surgical resections. (2) Methods: An in vitro model was based on two types of normal human melanocytes (HEMn-DP and HEMn-LP) and four melanoma cell lines (A375, WM1341D, Hs294T, and WM9). PARP1 mRNA gene expression was estimated using real-time polymerase chain reaction (RT-PCR), whereas the protein level of PARP1 was evaluated by fluorescence confocal microscopy and then confirmed by Western Blotting analysis. The expression of PARP1 was also assessed by immunohistochemistry in formalin-fixed paraffin-embedded tissues of 128 primary cutaneous melanoma patients and correlated with follow-up and clinicopathologic features. (3) Results: The in vitro study showed that melanoma cells exhibited significantly higher PARP1 expression at mRNA and protein levels than normal melanocytes. High PARP1 expression was also associated with the invasiveness of tumor cells. Elevated nuclear PARP1 expression in patients without nodal metastases strongly correlated with significantly shorter disease-free survival (p = 0.0015) and revealed a trend with shorter cancer-specific overall survival (p = 0.05). High PARP1 immunoreactivity in the lymph node-negative group of patients was significantly associated with higher Breslow tumor thickness, presence of ulceration, and a higher mitotic index (p = 0.0016, p = 0.023, and p < 0.001, respectively). In patients with nodal metastases, high PARP1 expression significantly correlated with the presence of microsatellitosis (p = 0.034), but we did not confirm the prognostic significance of PARP1 expression in these patients. In the entire analyzed group of patients (with and without nodal metastases at the time of diagnosis), PARP1 expression was associated with a high mitotic index (p = 0.001) and the presence of ulceration (p = 0.036). Moreover, in patients with elevated PARP1 expression, melanoma was more frequently located in the skin of the head and neck region (p = 0.015). In multivariate analysis, high PARP1 expression was an independent unfavorable prognosticator in lymph node-negative cutaneous melanoma patients. (4) Conclusions: In vitro molecular biology approaches demonstrated enhanced PARP1 expression in cutaneous melanoma. These results were confirmed by the immunohistochemical study with clinical parameter analysis, which showed that a high level of PARP1 correlated with unfavorable clinical outcome. These observations raise the potential role of PARP1 inhibitor-based therapy in cutaneous melanoma.

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Onconase Restores Cytotoxicity in Dabrafenib-Resistant A375 Human Melanoma Cells and Affects Cell Migration, Invasion and Colony Formation Capability

Cited by 14 publications

References 54 publications

Strengths and Challenges of Secretory Ribonucleases as AntiTumor Agents

Strengths and Challenges of Secretory Ribonucleases as AntiTumor Agents

RNase A Domain-Swapped Dimers Produced Through Different Methods: Structure–Catalytic Properties and Antitumor Activity

PARP1 as a Marker of an Aggressive Clinical Phenotype in Cutaneous Melanoma—A Clinical and an In Vitro Study

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