Actinomycin V Inhibits Migration and Invasion via Suppressing Snail/Slug-Mediated Epithelial-Mesenchymal Transition Progression in Human Breast Cancer MDA-MB-231 Cells In Vitro

Abstract

Actinomycin V, an analog of actinomycin D produced by the marine-derived actinomycete Streptomyces sp., possessing a 4-ketoproline instead of a 4-proline in actinomycin D. In this study, the involvement of snail/slug-mediated epithelial-mesenchymal transition (EMT) in the anti-migration and -invasion actions of actinomycin V was investigated in human breast cancer MDA-MB-231 cells in vitro. Cell proliferation effect was evaluated by 3-(4,5-Dimethylthiazol)-2,5-diphenyltetrazolium bromide (MTT) assay. Wound-healing and Transwell assay were performed to investigate the anti-migration and -invasion effects of actinomycin V. Western blotting was used to detect the expression levels of E-cadherin, N-cadherin, vimentin, snail, slug, zinc finger E-box binding homeobox 1 (ZEB1), and twist proteins and the mRNA levels were detected by rt-PCR. Actinomycin V showed stronger cytotoxic activity than that of actinomycin D. Actinomycin V up-regulated both of the protein and mRNA expression levels of E-cadherin and down-regulated that of N-cadherin and vimentin in the same cells. In this connection, actinomycin V decreased the snail and slug protein expression, and consequently inhibited cells EMT procession. Our results suggest that actinomycin V inhibits EMT-mediated migration and invasion via decreasing snail and slug expression, which exhibits therapeutic potential for the treatment of breast cancer and further toxicity investigation in vivo is needed.

The Novel Nature Microtubule Inhibitor Ivalin Induces G2/M Arrest and Apoptosis in Human Hepatocellular Carcinoma SMMC-7721 Cells In Vitro

“…We seeded the cells on glass over slips and treated them with 0 to 8 μmol/L Ivalin for 24 h. Immunofluorescence staining was performed as previously described [17].…”

“…We exposed the cells to indicated concentration of Ivalin for 24 h, then the protein expressions were analyzed by Western blotting which we carried out as previously described [17].…”

Involvement of Reactive Oxygen Species in the Hepatorenal Toxicity of Actinomycin V In Vitro and In Vivo

“…The cells were treated with serial concentrations of Act V and Act D (0–0.02 μM) for 48 h, and the IC 50 s were determined (Act D, which has been used for a long time in clinic, serves as a positive control here), as shown in Table 1 and our previous study [ 10 , 11 ]. Act V exhibited considerably stronger efficacy against cancer cells with a ten-fold lower IC 50 value compared to Act D. Act V and Act D induced cytotoxicity in human normal liver LO-2 and human embryonic kidney 293T cell lines in a concentration-dependent manner ( Figure 2 ).…”

“…[ 1 ], has a similar structure to that of Act D ( Figure 1 ). In our previous studies [ 10 , 11 ], Act V was found to be more cytotoxic to tumor cells than Act D. Moreover, Act V inhibited the migration and metastasis of breast cancer cells. In this aspect, Act V may be a potential substitute for Act D, so understanding the toxicological mechanisms of Act V and Act D is of great worth.…”

Direct cold atmospheric plasma and plasma‐activated medium effects on breast and cervix cancer cells

“…[53] Afterward, the absorption of samples was read at 570-nm wavelength using a microplate reader (model 680; Bio-Rad Laboratories, Hercules, CA). [54][55][56][57] 2.7 | Medium H 2 O 2 assay H 2 O 2 detection was done in the control group (without any treatment) after direct-plasma treatment on Hela cells and PAM treatments (5-min treatment with both methods [the best results] and with He + 0.5% O 2 [the best combination]) and the medium was immediately (<1 min) provided using a fluorimetric hydrogen peroxide assay kit (MAK165-1KT; Sigma-Aldrich) according to manufacturer's standard protocol. At each measurement, 50 μl of the medium was collected and transferred to a flat-bottomed 96-well plate.…”