Arsenic trioxide inhibits the growth of A498 renal cell carcinoma cells via cell cycle arrest or apoptosis

doi:10.1016/S0006-291X(02)02831-0

Biochemical and Biophysical Research Communications

Volume 300, Issue 1, 3 January 2003, Pages 230-235

https://doi.org/10.1016/S0006-291X(02)02831-0 Get rights and content

Abstract

Previously, we showed that arsenic trioxide potently inhibited the growth of myeloma cells and head and neck cancer cells. Here, we demonstrate that arsenic trioxide inhibited the proliferation of all the renal cell carcinoma cell lines (ACHN, A498, Caki-2, Cos-7, and Renca) except only one cell line (Caki-1) with IC₅₀ of about 2.5–10 μM. Arsenic trioxide induced a G₁ or a G₂-M phase arrest in these cells. When we examined the effects of this drug on A498 cells, arsenic trioxide (2.5 μM) decreased the levels of CDK2, CDK6, cyclin D1, cyclin E, and cyclin A proteins. Although p21 protein was not increased by arsenic trioxide, this drug markedly enhanced the binding of p21 with CDK2. In addition, the activities of CDK2- and CDK6-associated kinase were reduced in association with hypophosphorylation of Rb protein. Arsenic trioxide (10 μM) also induced apoptosis in A498 cells. Apoptotic process of A498 cells was associated with the changes of Bcl-_XL, caspase-9, caspase-3, and caspase-7 proteins as well as mitochondria transmembrane potential (Δψ_m) loss. Taken together, these results demonstrate that arsenic trioxide inhibits the growth of renal cell carcinoma cells via cell cycle arrest or apoptosis.

Section snippets

Materials and methods

Cell lines and culture. Renal cell carcinoma cell lines used in this study were ACHN, A498, Caki-1, Caki-2, Cos-7, and Renca. These cells were maintained according to the recommendation of the American Type Culture Collection. Cells were grown at 37 °C in an atmosphere of 5% CO₂ in air.

Reagent. Arsenic trioxide was purchased from Sigma Chemical (St. Louis, MO). Arsenic trioxide was dissolved in 1.65 M NaOH at 5×10⁻² M as a stock solution. The maximum concentration of NaOH in culture had no

Effect of arsenic trioxide on growth inhibition in renal cell carcinoma cell lines

We examined the effect of arsenic trioxide on the cell proliferation of renal cell carcinoma cell lines using MTT assay. Dose-dependent inhibition of cell growth was observed in all the cell lines but Caki-1 cells with IC₅₀ of about 2.5–10 μM following the treatment of arsenic trioxide for 72 h (Fig. 1). The susceptibility to arsenic trioxide in renal cell carcinoma cells is lower than that in leukemia and myeloma cell lines [2], [3], [4] and is similar to that in solid tumors such as

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Cited by (108)

Mechanism for arsenic-induced toxic effects
2023, Handbook of Arsenic Toxicology
The metalloid arsenic is a natural environmental contaminant to which humans are routinely exposed through food, water, air, and soil. It causes various biological effects on cells and tissues. The metabolism of arsenic has an important role in its toxicity. This metabolism involves methyltransferase mediated reduction to a trivalent state followed by oxidative methylation to a pentavalent state. The pentavalent arsenicals are reported to be less potent toxicants than the trivalent arsenicals including those that are methylated. Arsenic-induced toxicity is believed to be mediated via reactive oxygen species (ROS) generation, Ca^2 + accumulation, caspase-3 up-regulation, Bcl-2 down-regulation, and p-53 deficiency. Alterations of these factors from normal physiology might play prominent roles in carcinogenicity, cardiovascular, testicular, genotoxicity, diabetes, and nervous systems disorders. The exact mechanisms in arsenic toxicity and its subsequent organ pathophysiology are not well defined to date. Potential mechanisms of various types of arsenic-induced cancer may include oxidative stress, co-carcinogenesis and tumor promotion, genotoxicity, altered DNA methylation, altered cell proliferation, etc. As a preventive and curative measure of arsenic toxicity, combination therapies using antioxidants (N-acetylcysteine, α-lipoic acid, quercetin, etc.) and a thiol-chelating agent (with a proper chelator) can be considered as a better therapeutic approach than chelation therapy alone. We intend to summarize the up-to-date knowledge from literature on the molecular and cellular mechanisms of arsenic toxicity and usefulness of antioxidants in preventing these alterations.
Different mechanisms of arsenic related signaling in cellular proliferation, apoptosis and neo-plastic transformation
2021, Ecotoxicology and Environmental Safety
Citation Excerpt :
When arsenic breaks DNA strands, p53 accumulation occurs as a sequel of p53 activation by the enhanced levels of mdm2 (murine double minute-2) and p21. It ensures the cell-cycle arrest at G2M phase (Park et al., 2003). In arsenic related induction process, assessment of phosphorylation in serine 15 site of p53 showed that this site is specific for phosphorylation by ATM which is a member of PI3 kinase-allied protein kinase (Yih and Lee, 2000).
Arsenic is a toxic heavy metal vastly dispersed all over the earth crust. It manifests several major adverse health issues to millions of arsenic exposed populations. Arsenic is associated with different types of cancer, cardiovascular disorders, diabetes, hypertension and many other diseases. On the contrary, arsenic (arsenic trioxide, As₂O₃) is used as a chemotherapeutic agent in the treatment of acute promyelocytic leukemia. Balance between arsenic induced cellular proliferations and apoptosis finally decide the outcome of its transformation rate. Arsenic propagates signals via cellular and nuclear pathways depending upon the chemical nature, and metabolic-fates of the arsenical compounds. Arsenic toxicity is propagated via ROS induced stress to DNA-repair mechanism and mitochondrial stability in the cell. ROS induced alteration in p53 regulation and some mitogen/ oncogenic functions determine the transformation outcome influencing cyclin-cdk complexes. Growth factor regulator proteins such as c-Jun, c-fos and c-myc are influenced by chronic arsenic exposure. In this review we have delineated arsenic induced ROS regulations of epidermal growth factor receptor (EGFR), NF-ĸβ, MAP kinase, matrix-metalloproteinases (MMPs). The role of these signaling molecules has been discussed in relation to cellular apoptosis, cellular proliferation and neoplastic transformation. The arsenic stimulated pathways which help in proliferation and neoplastic transformation ultimately resulted in cancer manifestation whereas apoptotic pathways inhibited carcinogenesis. Therapeutic strategies against arsenic should be designed taking into account all these factors.
Counteracting arsenic toxicity: Curcumin to the rescue?
2020, Journal of Hazardous Materials
Citation Excerpt :
As can damage the DNA and triggers the cell arrest at G or G/M phase (Chen and Shi, 2002; Hossain et al., 2000). Arsenic trioxide (As2O3) can attenuate the steady-state levels of CDK/cyclin complexes, such as CDK2, CDK6, cyclins A, D1 and E, in different tumor cells (Park et al., 2000, 2003). As2O3 can arrest the cell cycle by the suppression of cdc2-associated kinase and CDK2/6-associated kinases in compliance with the reduction of cdc25B/C phosphatases and the Rb protein hypophosphorylation, respectively (Park et al., 2000, 2003).
Arsenicosis leads to various irreversible damages in several organs and is considered to be a carcinogen. The effects of chronic arsenic poisoning are a result of an imbalance between pro- and antioxidant homeostasis, oxidative stress, as well as DNA and protein damage. Curcumin, the polyphenolic pigment extracted from the rhizome of Curcuma longa, is well-known for its pleiotropic medicinal effects. Curcumin has been shown to have ameliorative effects in arsenic-induced genotoxicity, nephrotoxicity, hepatotoxicity, angiogenesis, skin diseases, reproductive toxicity, neurotoxicity, and immunotoxicity. This review aims to summarize the scientific evidence on arsenic toxicity in various organs and the ameliorative effects of curcumin on the arsenic toxicity.
Arsenic trioxide disturbs the LIS1/NDEL1/dynein microtubule dynamic complex by disrupting the CLIP170 zinc finger in head and neck cancer
2020, Toxicology and Applied Pharmacology
Cancer mortality is mainly caused by metastasis, which requires dynamic remodeling of cytoskeletal components such as microtubules. Targeting microtubules presents a promising antimetastatic strategy that could prevent cancer spreading and recurrence. It is known that arsenic trioxide (ATO) is able to inhibit the migration and invasion of solid malignant tumors, but its exact molecular mechanism remains unclear. Here, we report a novel molecular target and antimetastatic mechanism of ATO in head and neck squamous cell carcinoma (HNSCC). We found that cytoplasmic linker protein 170 (CLIP170) was overexpressed in HNSCC tissues and cells compared to normal controls. ATO at non-cytotoxic level (1 μM) inhibited the migration and invasion of HNSCC cells by displacing zinc in the zinc finger motif of CLIP170, which is a key protein that controls microtubule dynamics. The antimetastatic effects of ATO were equivalent to those of siRNA-mediated CLIP170 knockdown. Furthermore, ATO dysregulated microtubule polymerization via the CLIP170/LIS1/NDEL1/dynein signaling pathway in Cal27 cells as a functional consequence of CLIP170 zinc finger disruption. The effect was partially reversed by zinc supplementation. Taken together, these findings reveal that CLIP170 is a novel molecular target of ATO and demonstrate the capability and underlying mechanisms of ATO as a potential antimetastatic agent for HNSCC treatment.
Surface-modified PLGA nanoparticles with PEG/LA-chitosan for targeted delivery of arsenic trioxide for liver cancer treatment: Inhibition effects enhanced and side effects reduced
2019, Colloids and Surfaces B: Biointerfaces
Arsenic trioxide (As₂O₃), an effective drug for leukemia, is limited to be used for solid tumor treatment due to its high side effects. In this study, polyethylene glycol (PEG) and lactobionic acid (LA) modified chitosan (PLC) was synthesized and was used to coat poly(lactide-co-glycolide) (PLGA) nanoparticles for encapsulation and targeted release of As₂O₃ in liver cancer treatment. The As₂O₃-loaded PLGA/PLC nanoparticles (As₂O₃-PLGA/PLC NPs) were fabricated through double emulsion-solvent evaporation method and were optimized by orthogonal tests. As₂O₃-PLGA/PLC NPs presented suitable physical stability, positive charge, high encapsulation efficiency and drug loading, and good biocompatibility. As expected, the NPs can quickly release enough dose of As₂O₃ in a short time and then sustain the drug concentration. The As₂O₃-PLGA/PLC NPs showed effective inhibition of SMMC-7721 cells while having lower cytotoxicity against normal human liver cells (LO2 cells). Furthermore, In vivo study showed that the NPs did not present toxic effects on kidney and liver, but showed relatively high growth inhibition effect on liver tumor. Therefore, this PLGA/PLC NPs could be an effective and safe drug delivery system for liver cancer chemotherapy.
Evaluation of phyto-medicinal efficacy of thymoquinone against Arsenic induced mitochondrial dysfunction and cytotoxicity in SH-SY5Y cells
2019, Phytomedicine
It is evaluated that a few million individuals worldwide are experiencing Arsenic (As) harmfulness coming about because of anthropogenic discharges. There is likewise proof to propose that As can affect the peripheral, as well as, the central nervous system (CNS). On the contrary, thymoquinone (TQ), a biologically active ingredient of Nigella sativa has exhibited numerous neuro-pharmacological traits since ancient times.
In the present study, the neuroprotective efficacy of TQ was explored by primarily studying its antioxidant and anti-apoptotic potential against Arsenic trioxide (As₂O₃) induced toxicity in SH-SY5Y human neuroblastoma cell lines.
For experimentation, cells were seeded in 96 well tissue culture plates and kept undisturbed for 24 h to attain proper adhesion. After 75–80% confluence, cells were pretreated with 10 µM and 20 µM thymoquinone (TQ) for 1 h After adding 2 µM As, cells were set aside for incubation for 24 h without changing the medium.
The mitigatory effects of TQ with particular reference to cell viability and cytotoxicity, the generation of reactive oxygen species, DNA damage, and mitochondrial dynamics were studied.
Pretreatment of SH-SY5Y cells with TQ (10 and 20 μM) for an hour and subsequent exposure to 2 μM As₂O₃ protected the SH-SY5Y cells against the neuro-damaging effects of the latter. Also, the SH-SY5Y cells were better preserved with increased viability, repaired DNA, less free radical generation and balanced transmembrane potential than those exposed to As₂O₃ alone. TQ pretreatment also inhibited As₂O₃-induced exacerbation in protein levels of BAX and PARP-1 and restored the loss of Bcl₂ levels.
The findings of this study suggest that TQ may prevent neurotoxicity and As₂O₃-induced apoptosis and cytotoxicity. It is, therefore, worth studying further for its potential to reduce the risks of arsenic-related neurological implications.

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Arsenic trioxide inhibits the growth of A498 renal cell carcinoma cells via cell cycle arrest or apoptosis

Abstract

Section snippets

Materials and methods

Effect of arsenic trioxide on growth inhibition in renal cell carcinoma cell lines

Blood

Blood

Biochem. Biophys. Res. Commun.

Pharmacol. Ther.

Cell

Curr. Opin. Cell Biol.

Cell

Pharmacol. Ther.

Leukemia Res.

FEBS Lett.

Toxicol. Appl. Pharmacol.

Complete remission after treatment of acute promyelocytic leukemia with arsenic trioxide

N. Engl. J. Med.

Arsenic trioxide as an inducer of apoptosis and loss of PML/RARα protein in acute promyelocytic leukemia cells

J. Natl. Cancer Inst.

Arsenic trioxide-mediated growth inhibition in MC/CAR myeloma cells via cell cycle arrest in association with induction of cyclin-dependent kinase inhibitor, p21, and apoptosis

Cancer Res.

The induction of apoptosis and cell cycle arrest by arsenic trioxide in lymphoid neoplasms

Leukemia