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Home>>Signaling Pathways>> Ubiquitination/ Proteasome>> Autophagy>>Actinomycin D

Actinomycin D (Synonyms: Cosmegen, Dactinomycin, Meractinomycin, NCI C04682, NSC 3053, Oncostatin K)

Catalog No.GC16866 Copy One-Click Copy Product Info

Actinomycin D is a type of actinomycin cyclic peptide isolated from Streptomyces fermentation products. Actinomycin D can inhibit DNA repair (IC50=0.42μM) and mRNA synthesis, and is commonly used to study RNA stability.

Products are for research use only. Not for human use. We do not sell to patients.

Actinomycin D Chemical Structure

Cas No.: 50-76-0

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1mg
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10mg
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25mg
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50mg
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100mg
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Sample solution is provided at 25 µL, 10mM.



Product has been cited by 121 publications

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Description Protocol Chemical Properties Product Documents Related Video Related Products

Description of Actinomycin D

Actinomycin D is a type of actinomycin cyclic peptide isolated from Streptomyces fermentation products. Actinomycin D can inhibit DNA repair (IC50=0.42μM) and mRNA synthesis, and is commonly used to study RNA stability. Actinomycin D intercalates into GC-rich regions of DNA duplex to block RNA polymerase elongation, thereby inhibiting transcription (effective in both eukaryotes and prokaryotes). Actinomycin D exerts anti-tumor effects by inducing DNA damage, cell cycle arrest and apoptosis. Actinomycin D can be used in research related to childhood malignancies such as Ewing sarcoma and Wilms tumor, as well as antiviral (including COVID-19) studies[1-4].

In vitro, treatment of human chronic myeloid leukemia K562 cells with Actinomycin D (0.04-4μM) combined with salvicine (6.25μM) for 24 hours resulted in Actinomycin D concentration-dependently inhibiting salvicine-induced apoptosis[5]. Treatment of A549, NCI-H28, U-2 OS, NCI-H292, A375 and other multi-origin cancer cell lines with Actinomycin D (5nM) for 48 hours upregulated p53 and phosphorylated p53 Ser15/Ser33/Ser37/Ser46, induced p21 and MDM2 expression, and downregulated the mTOR activity marker p-p70S6K(Thr389)[6]. Treatment of SK-N-JD, SH-SY5Y (p53 wild-type) and LA1-55n, SK-N-AS (p53 deleted/truncated) neuroblastoma cells with Actinomycin D (0.1-10nM) for 24-48 hours reduced cell viability[7].

In vivo, intraperitoneal injection of Actinomycin D (0.5mg/kg; single injection) into Swiss mice 60 minutes before dexamethasone administration blocked the inhibitory effect of dexamethasone on mouse locomotion[8]. Intratumoral injection of Actinomycin D (1μg; 7 consecutive days) combined with tumor necrosis factor α (5μg) into B6D2F1 or BALB/c mice bearing MmB16 melanoma, LL/2 lung carcinoma or L1 sarcoma significantly delayed tumor growth[9]. A single intraperitoneal injection of Actinomycin D (0.5mg/kg) given 24 hours before to 24 hours after irradiation did not aggravate lip mucosal radiation damage in NMRI mice, nor did Actinomycin D affect the rate of sublethal damage repair. Intraperitoneal injection of Actinomycin D (0.15mg/kg; once daily) on days 3-7 after irradiation (for 5 days) inhibited lip mucosal repopulation and enhanced late radiation damage in NMRI mice[10].

References:

[1] Barret JM, Salles B, Provot C, et al. Evaluation of DNA repair inhibition by antitumor or antibiotic drugs using a chemiluminescence microplate assay. Carcinogenesis. 1997 Dec;18(12):2441-5.

[2] Wu CH, Pan JS, Chang WC, et al. The molecular mechanism of actinomycin D in preventing neointimal formation in rat carotid arteries after balloon injury. J Biomed Sci. 2005;12(3):503-12.

[3] Merkel O, Wacht N, Sifft E, et al. Actinomycin D induces p53-independent cell death and prolongs survival in high-risk chronic lymphocytic leukemia. Leukemia. 2012 Dec;26(12):2508-16.

[4] Kleeff J, Kornmann M, Sawhney H, et al. Actinomycin D induces apoptosis and inhibits growth of pancreatic cancer cells. Int J Cancer. 2000 May 1;86(3):399-407.

[5] Qing C, Miao ZH, Tong LJ, et al. Actinomycin D inhibiting K562 cell apoptosis elicited by salvicine but not decreasing its cytotoxicity. Acta Pharmacol Sin. 2003 May;24(5):415-21.

[6] Zajkowicz A, Gdowicz-Kłosok A, Krześniak M, et al. Actinomycin D and nutlin-3a synergistically promote phosphorylation of p53 on serine 46 in cancer cell lines of different origin. Cell Signal. 2015 Jul;27(7):1444-54.

[7] Cortes CL, Veiga SR, Almacellas E, et al. Effect of low doses of actinomycin D on neuroblastoma cell lines. Mol Cancer. 2016 Jan 4;15:1.

[8] Capasso A, Di Giannuario A, Loizzo A, et al. Actinomycin D blocks the reducing effect of dexamethasone on amphetamine and cocaine hypermotility in mice. Gen Pharmacol. 1996;27(4):707-12.

[9] Lasek W, Giermasz A, Kuc K, et al. Potentiation of the anti-tumor effect of actinomycin D by tumor necrosis factor α in mice: correlation between in vitro and in vivo results. Int J Cancer. 1996;66(3):374-9.

[10] Landuyt W, Van der Schueren E, Ang KK. The effect of actinomycin D on radiation induced reactions of the lip mucosa of mice. Int J Radiat Oncol Biol Phys. 1985;11(8):1503-12.

Protocol of Actinomycin D

Experimental Protocol for Measuring mRNA Stability Using Actinomycin D[1]

This experimental protocol is compiled based on research content and is for reference only. Adjust according to actual needs.

(1) Reagent preparation: Dissolve Actinomycin D in dimethyl sulfoxide (DMSO) to prepare a 1mg/mL stock solution. Aliquot and store at -20°C. Prepare cell culture medium. Prepare 0.25% trypsin, phosphate-buffered saline (PBS), TRIzol NC Reagent (Catalog No.GK20010), isopropanol, 75% ethanol, RNase-free water, etc.

(2) Cell culture and sample collection: Collect cells from one well of a 6-well plate using 0.1% trypsin at 37°C for 3min or using a cell scraper. Centrifuge at 470g for 3min. Discard the supernatant. Resuspend the cell pellet in 0.5mL of TRIzol NC Reagent (Catalog No.GK20010) (t=0) and freeze at -80°C. Add 30μL of 1mg/mL Actinomycin D stock solution (final concentration 10μg/mL) to each of the remaining five wells. Mix gently. Collect cells at 1, 2, 4, 6, and 8 hours after Actinomycin D addition. Freeze each cell pellet in 1mL of TRIzol NC Reagent (Catalog No.GK20010).

(3) RNA extraction: Extract RNA from samples following the instructions for GK20010 (Catalog No.GK20010). Briefly air-dry the RNA pellet for 5-10min. Avoid over-drying. Dissolve RNA in 30μL of RNase-free water.

(4) Reverse transcription and quantitative real-time PCR: Perform DNase I treatment on all RNA samples. It is recommended to use SuperFast RT Master Mix for qPCR (gDNA remover) (Catalog No.GK10031) for reverse transcription. Dilute the obtained cDNA 10-fold with RNase-free water as the template for qPCR. Use SYBR Green qPCR Master Mix (Catalog No.GK20002) for quantitative real-time PCR. Perform at least two technical replicates for each sample.

(5) Data analysis: After qPCR completion, calculate the average Ct value of replicates for each time point. Use the average Ct at t=0 as the reference. Calculate ΔCt = (average Ct of each time point – average Ct of t=0). Calculate relative mRNA abundance = 2(-ΔCt).

Precautions:

(1) Prepare Actinomycin D stock solution in DMSO and store in aliquots at -20°C protected from light. Avoid repeated freeze-thaw cycles.

(2) For different cell types, optimize the Actinomycin D concentration.

(3) Do not air-dry the RNA pellet for more than 10min. Over-drying significantly decreases RNA solubility.

(4) Include melt curve analysis in qPCR to confirm amplification of a single specific product (single peak).

(5) For safety and health, wear a lab coat and disposable gloves during operation.

References:

[1] Ratnadiwakara M, Änkö ML. mRNA Stability Assay Using transcription inhibition by Actinomycin D in Mouse Pluripotent Stem Cells. Bio Protoc. 2018 Nov 5;8(21):e3072.

Chemical Properties of Actinomycin D

Cas No. 50-76-0 SDF
Synonyms Cosmegen, Dactinomycin, Meractinomycin, NCI C04682, NSC 3053, Oncostatin K
Chemical Name 2-amino-4,6-dimethyl-3-oxo-1-N,9-N-bis[7,11,14-trimethyl-2,5,9,12,15-pentaoxo-3,10-di(propan-2-yl)-8-oxa-1,4,11,14-tetrazabicyclo[14.3.0]nonadecan-6-yl]phenoxazine-1,9-dicarboxamide
Canonical SMILES CC1C(C(=O)NC(C(=O)N2CCCC2C(=O)N(CC(=O)N(C(C(=O)O1)C(C)C)C)C)C(C)C)NC(=O)C3=C4C(=C(C=C3)C)OC5=C(C(=O)C(=C(C5=N4)C(=O)NC6C(OC(=O)C(N(C(=O)CN(C(=O)C7CCCN7C(=O)C(NC6=O)C(C)C)C)C)C(C)C)C)N)C
Formula C62H86N12O16 M.Wt 1255.43
Solubility ≥ 62.75 mg/mL in DMSO Storage Store at -20℃,protected from light
General tips Please select the appropriate solvent to prepare the stock solution according to the solubility of the product in different solvents; once the solution is prepared, please store it in separate packages to avoid product failure caused by repeated freezing and thawing.Storage method and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored at -20°C, please use it within 1 month.
To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time.
Shipping Condition Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request.

Complete Stock Solution Preparation Table of Actinomycin D

Prepare stock solution
1 mg 5 mg 10 mg
1 mM 796.5 μL 3.9827 mL 7.9654 mL
5 mM 159.3 μL 796.5 μL 1.5931 mL
10 mM 79.7 μL 398.3 μL 796.5 μL

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In vivo Formulation Calculator (Clear solution) of Actinomycin D

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    Actinomycin D-GlpBio

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Average Rating: 5 ★★★★★ (Based on Reviews and 30 reference(s) in Google Scholar.)

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