Withaferin-A Inhibits Growth of Drug-Resistant Breast Carcinoma by Inducing Apoptosis and Autophagy, Endogenous Reactive Oxygen Species (ROS) Production, and Inhibition of Cell Migration and Nuclear Factor kappa B (Nf-Œ∫B)/Mammalian Target of Rapamycin (m-TOR) Signalling Pathway
Authors: Yanwei Wang
Yu Li
Xiaokang Liu
Qiang Ma
Ai Lin Song
Journal: Medical Science Monitor (2019)
Discipline: CLINICAL MEDICINE
Keywords: withaferin-A; breast carcinoma cells; anticancer effect; autophagy; cell apoptosis
Affiliations: Fifth Department of General Surgery, Second Hospital of Lanzhou University, Lanzhou, Gansu, China (mainland).

Abstract:  BACKGROUND The main purpose of this study was to assess in vitro and in vivo the anticancer effect of withaferin-A in human breast carcinoma cells (MDA-MB-231), and to assess its effects on autophagy, cell apoptosis, ROS production, cell migration and invasion, and Nf-kappaB/m-TOR signalling pathway. MATERIAL AND METHODS Proliferation of MDA-MB-231 cells at various doses of the drug was studied by CCK8 cell viability assay. Effects on cell apoptosis were studied by fluorescence microscopy in combination with flow cytometry and Western blot analysis. Effects on autophagy were evaluated by transmission electron microscopy and Western blot. Effects on cellular migration were examined in vitro by wound healing assay. RESULTS The results indicated that withaferin-A led to significant reduction of MDA-MB-231 cell viability. The anticancer action of withaferin-A was shown to be due to the stimulation of autophagy, which was accompanied by enhancement of LC3 expression. Withaferin-A prompted mitochondrial apoptosis, which was also associated with increased level of Bax and decreased Bcl-2 in MDA-MB-231 cells. It was also observed that withaferin-A has decreases cellular migration and invasion of the tested human breast cancer cells. The effects of withaferin-A were also investigated in vivo, and it was found that this molecule could inhibit the growth of tumor xenografts in tested mice. Withaferin-A led to suppression of the Nf-kappaB/m-TOR signalling pathway. CONCLUSIONS In brief, the withaferin-A molecule has great potential as an anticancer agent against drug-resistant breast cancer, and as such needs to be further studied in detail. 

Retracted: Yes

PubPeer comments:
 *[right]* Figure 3 from [Liu et al (2019)] . Electron microscopy images of withaferin-A-treated MDA-MB-231 cells showing formation of autophagosomes, thus indicating induction of autophagy. *[left]* Figure 7 from [Chu et al (2020)] .  Electron microscopy showing that Daidzein induces autophagy in the A-375 cells at indicated concentrations (arrows depict the autophagosomes).  , 
 *[left]* Figure 10 from [Liu et al (2019)] .  Western blot analysis showing concentration-dependent inhibition of the Nf-kB m-TOR expression in human breast cancer cells upon treatment with varied concentrations of withaferin-A. *[right]* Figure 8 from [Zhang et al (2019)] .  Western blot analysis showing concentration-dependent inhibition of the NF-kB expression in HeLa cells upon treatment with varied concentrations of AL.  , 
 *[left]* Figure 6 from [Liu et al (2019)] .  Annexin V PI assay showing percentage of apoptotic MDA-MB-231 cells at indicated concentrations of withaferin-A. *[right]* Fig. 2b from [Li et al (2020)] .  Annexin V PI staining showing the percentage of apoptotic U87 and U118 cells .  Figure 3B from [Xu et al (2019)] .  miR-181 induces apoptosis in glioma cells. ...  (B) annexin V PI staining of miR-NC and miR-181 mimics transfected U87 and U118 cells showing induction of apoptosis.  UPDATE: Another sighting! Figs 1E, 3H from [Zhang et al (2020)] .  Suppression of miR-1179 inhibits proliferation of ovarian cancer cell lines:... annexin V PI staining of NC or miR-1179 inhibitor transfected PA-1 and SK-OV-3 ;  miR-1179 exerts its effects by targeting PTEN... Annexin V PI staining of NC or pcDNA-PTEN transfected PA-1 ovarian cancer cells  , 
 Retracted [25 March 2021] .  Retracted, due to breach of publishing guidelines, following the identification of non-original and manipulated figure images. ,