scholarly journals Annurca apple polyphenol extract selectively kills MDA-MB-231 cells through ROS generation, sustained JNK activation and cell growth and survival inhibition

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Elisa Martino ◽  
Daniela Cristina Vuoso ◽  
Stefania D’Angelo ◽  
Luigi Mele ◽  
Nunzia D’Onofrio ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Triple Negative Breast Cancer ◽  
Triple Negative ◽  
Specific Inhibitor ◽  
Underlying Mechanism ◽  
Ros Generation ◽  
Growth And Survival ◽  
Phase Arrest ◽  
M Phase ◽  
Jnk Activation

Abstract Polyphenols represent the most studied class of nutraceuticals that can be therapeutics for a large spectrum of diseases, including cancer. In this study, we investigated for the first time the antitumor activities of polyphenol extract from Annurca apple (APE) in MDA-MB-231 triple negative breast cancer cells, and we explored the underlying mechanisms. APE selectively inhibited MDA-MB-231 cell viability and caused G2/M phase arrest associated with p27 and phospho-cdc25C upregulation and with p21 downregulation. APE promoted reactive oxygen species (ROS) generation in MDA-MB-231 cells while it acted as antioxidant in non-tumorigenic MCF10A cells. We demonstrated that ROS generation represented the primary step of APE antitumor activity as pretreatment with antioxidant N-acetylcysteine (NAC) prevented APE-induced G2/M phase arrest, apoptosis, and autophagy. APE downregulated Dusp-1 and induced a significant increase in JNK/c-Jun phosphorylation that were both prevented by NAC. Moreover, downregulation of JNK by its specific inhibitor SP600125 significantly diminished the anticancer activity of APE indicating that ROS generation and sustained JNK activation represented the main underlying mechanism of APE-induced cell death. APE also inhibited AKT activation and downregulated several oncoproteins, such as NF-kB, c-myc, and β-catenin. In light of these results, APE may be an attractive candidate for drug development against triple negative breast cancer.

scholarly journals Cucurbitacin E Induces Cell Cycle G2/M Phase Arrest and Apoptosis in Triple Negative Breast Cancer

PLoS ONE ◽  
2014 ◽  
Vol 9 (7) ◽  
pp. e103760 ◽  
Author(s):  
Yanjie Kong ◽  
Jianchao Chen ◽  
Zhongmei Zhou ◽  
Houjun Xia ◽  
Ming-Hua Qiu ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cell Cycle ◽  
Triple Negative Breast Cancer ◽  
Triple Negative ◽  
Phase Arrest ◽  
M Phase ◽  
Cucurbitacin E

scholarly journals Anticancer Effects and Molecular Action of 7-α-Hydroxyfrullanolide in G2/M-Phase Arrest and Apoptosis in Triple Negative Breast Cancer Cells

Molecules ◽  
2022 ◽  
Vol 27 (2) ◽  
pp. 407
Author(s):  
Siriphorn Chimplee ◽  
Sittiruk Roytrakul ◽  
Suchada Sukrong ◽  
Theera Srisawat ◽  
Potchanapond Graidist ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Triple Negative Breast Cancer ◽  
Mechanism Of Action ◽  
Breast Cancer Cells ◽  
Low Dose ◽  
Triple Negative ◽  
Phase Arrest ◽  
M Phase ◽  
Caspase 7 ◽  
Molecular Mechanism Of Action

Triple negative breast cancer (TNBC) is a breast cancer subtype characterized by the absence of estrogen receptor, progesterone receptor and human epidermal growth factor receptor 2 expression. TNBC cells respond poorly to targeted chemotherapies currently in use and the mortality rate of TNBC remains high. Therefore, it is necessary to identify new chemotherapeutic agents for TNBC. In this study, the anti-cancer effects of 7-α-hydroxyfrullanolide (7HF), derived from Grangea maderaspatana, on MCF-7, MDA-MB-231 and MDA-MB-468 breast cancer cells were assessed using MTT assay. The mode of action of 7HF in TNBC cells treated with 6, 12 and 24 µM of 7HF was determined by flow cytometry and propidium iodide (PI) staining for cell cycle analysis and annexin V/fluorescein isothiocyanate + PI staining for detecting apoptosis. The molecular mechanism of action of 7HF in TNBC cells was investigated by evaluating protein expression using proteomic techniques and western blotting. Subsequently, 7HF exhibited the strongest anti-TNBC activity toward MDA-MB-468 cells and a concomitantly weak toxicity toward normal breast cells. The molecular mechanism of action of low-dose 7HF in TNBC cells primarily involved G2/M-phase arrest through upregulation of the expression of Bub3, cyclin B1, phosphorylated Cdk1 (Tyr 15) and p53-independent p21. Contrastingly, the upregulation of PP2A-A subunit expression may have modulated the suppression of various cell survival proteins such as p-Akt (Ser 473), FoxO3a and β-catenin. The concurrent apoptotic effect of 7HF on the treated cells was mediated via both intrinsic and extrinsic modes through the upregulation of Bax and active cleaved caspase-7–9 expression and downregulation of Bcl-2 and full-length caspase-7–9 expression. Notably, the proteomic approach revealed the upregulation of the expression of pivotal protein clusters associated with G1/S-phase arrest, G2/M-phase transition and apoptosis. Thus, 7HF exhibits promising anti-TNBC activity and at a low dose, it modulates signal transduction associated with G2/M-phase arrest and apoptosis.

scholarly journals GCH1 induces immunosuppression through metabolic reprogramming and IDO1 upregulation in triple-negative breast cancer

2021 ◽  
Vol 9 (7) ◽  
pp. e002383
Author(s):  
Jin-Li Wei ◽  
Si-Yu Wu ◽  
Yun-Song Yang ◽  
Yi Xiao ◽  
Xi Jin ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Triple Negative Breast Cancer ◽  
Triple Negative ◽  
Molecular Taxonomy ◽  
Underlying Mechanism ◽  
Metabolic Reprogramming ◽  
Sequencing Data ◽  
Aryl Hydrocarbon

PurposeRegulatory T cells (Tregs) heavily infiltrate triple-negative breast cancer (TNBC), and their accumulation is affected by the metabolic reprogramming in cancer cells. In the present study, we sought to identify cancer cell-intrinsic metabolic modulators correlating with Tregs infiltration in TNBC.Experimental designUsing the RNA-sequencing data from our institute (n=360) and the Molecular Taxonomy of Breast Cancer International Consortium TNBC cohort (n=320), we calculated the abundance of Tregs in each sample and evaluated the correlation between gene expression levels and Tregs infiltration. Then, in vivo and in vitro experiments were performed to verify the correlation and explore the underlying mechanism.ResultsWe revealed that GTP cyclohydrolase 1 (GCH1) expression was positively correlated with Tregs infiltration and high GCH1 expression was associated with reduced overall survival in TNBC. In vivo and in vitro experiments showed that GCH1 increased Tregs infiltration, decreased apoptosis, and elevated the programmed cell death-1 (PD-1)-positive fraction. Metabolomics analysis indicated that GCH1 overexpression reprogrammed tryptophan metabolism, resulting in L-5-hydroxytryptophan (5-HTP) accumulation in the cytoplasm accompanied by kynurenine accumulation and tryptophan reduction in the supernatant. Subsequently, aryl hydrocarbon receptor, activated by 5-HTP, bound to the promoter of indoleamine 2,3-dioxygenase 1 (IDO1) and thus enhanced the transcription of IDO1. Furthermore, the inhibition of GCH1 by 2,4-diamino-6-hydroxypyrimidine (DAHP) decreased IDO1 expression, attenuated tumor growth, and enhanced the tumor response to PD-1 blockade immunotherapy.ConclusionsTumor-cell-intrinsic GCH1 induced immunosuppression through metabolic reprogramming and IDO1 upregulation in TNBC. Inhibition of GCH1 by DAHP serves as a potential immunometabolic strategy in TNBC.

scholarly journals Annurca apple polyphenol extract promotes mesenchymal-to-epithelial transition and inhibits migration in triple-negative breast cancer cells through ROS/JNK signaling

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Daniela Cristina Vuoso ◽  
Stefania D’Angelo ◽  
Rosalia Ferraro ◽  
Sergio Caserta ◽  
Stefano Guido ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Triple Negative Breast Cancer ◽  
Triple Negative ◽  
Live Cells ◽  
Ros Generation ◽  
Filamentous Actin ◽  
Microscopy Imaging ◽  
Jnk Signaling ◽  
Mesenchymal Transition ◽  
E Cadherin

Abstract Aberrant activation of epithelial-to-mesenchymal transition has been shown to correlate with triple-negative breast cancer (TNBC) progression and metastasis. Thus, the induction of the reverse process might offer promising opportunities to restrain TNBC metastatic spreading and related mortality. Recently, the Annurca apple polyphenol extract (APE) has been highlighted as a multi-faceted agent that selectively kills TNBC cells by ROS generation and sustained JNK activation. Here, by qualitatively and quantitatively monitoring the real-time movements of live cells we provided the first evidence that APE inhibited the migration of MDA-MB-231 and MDA-MB-468 TNBC cells and downregulated metalloproteinase-2 and metalloproteinase-9. In MDA-MB-231 cells APE decreased SMAD-2/3 and p-SMAD-2/3 levels, increased E-cadherin/N-cadherin protein ratio, induced the switch from N-cadherin to E-cadherin expression and greatly reduced vimentin levels. Confocal and scanning electron microscopy imaging of APE-treated MDA-MB-231 cells evidenced a significant cytoskeletal vimentin and filamentous actin reorganization and revealed considerable changes in cell morphology highlighting an evident transition from the mesenchymal to epithelial phenotype with decreased migratory features. Notably, all these events were reverted by N-acetyl-l-cysteine and JNK inhibitor SP600125 furnishing evidence that APE exerted its effects through the activation of ROS/JNK signaling. The overall data highlighted APE as a potential preventing agent for TNBC metastasis.

scholarly journals The testis protein ZNF165 is a SMAD3 cofactor that coordinates oncogenic TGFβ signaling in triple-negative breast cancer

2020 ◽  
Author(s):  
Zane A. Gibbs ◽  
Luis C. Reza ◽  
Chun-Chun Cheng ◽  
Jill M. Westcott ◽  
Kathleen McGlynn ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Triple Negative Breast Cancer ◽  
Triple Negative ◽  
Transforming Growth Factor ◽  
Zinc Finger Protein ◽  
Transforming Growth Factor Β ◽  
Aberrant Expression ◽  
Growth And Survival ◽  
Ct Antigens

ABSTRACTCancer/testis (CT) antigens are proteins whose expression is normally restricted to germ cells yet aberrantly activated in tumors, where their functions remain relatively cryptic. Here we report that ZNF165, a CT antigen frequently expressed in triple-negative breast cancer (TNBC), associates with SMAD3 to modulate transcription of transforming growth factor β (TGFβ)-dependent genes and thereby promote growth and survival. In addition, we identify the KRAB zinc finger protein, ZNF446, and its associated tripartite motif protein, TRIM27, as obligate components of the ZNF165-SMAD3 complex that also support tumor cell viability. Importantly, we find that TRIM27 alone is necessary for ZNF165 transcriptional activity and is required for orthotopic tumor growth in vivo. Our findings indicate that aberrant expression of a testis-specific transcription factor is sufficient to co-opt somatic transcriptional machinery to drive a pro-tumorigenic gene expression program.

scholarly journals SphK2/S1P promotes metastasis in triple negative breast cancer through PAK1/LIMK1/Cofilin1 signaling pathway

2020 ◽  
Author(s):  
Weiwei Shi ◽  
Ding Ma ◽  
Yin Cao ◽  
Lili Hu ◽  
Shuwen Liu ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Antitumor Activity ◽  
Signaling Pathway ◽  
Lung Metastasis ◽  
Triple Negative Breast Cancer ◽  
Molecular Mechanisms ◽  
Triple Negative ◽  
Specific Inhibitor ◽  
Downstream Signaling

Abstract Background: Triple negative breast cancer (TNBC) features poor prognosis which partialy attributed to the high metastasis rate. However, there is no effective target for systemic TNBC therapy due to the absence of estrogen, progesterone, and human epidermal growth factor 2 receptors (ER, PR, HER-2) up to date. In the present study, we evaluated the role of sphingosine kinase 2 (SphK2) and its catalysate sphingosine-1-phosphate (S1P) in TNBC metastasis, and the antitumor activity of SphK2 specific inhibitor ABC294640 in TNBC metastasis. Methods: The function of SphK2 and S1P in migration of TNBC cells was evaluated by Transwell migration and wound healing assays. The molecular mechanisms of SphK2/S1P mediating TNBC metastasis were investigated using cell line establishment, western blot, histological examination and immunohistochemistry assays. The antitumor activity of ABC294640 was examined in TNBC lung metastasis model in vivo. Results: SphK2 regulated TNBC cells migration through the generation of S1P. Targeting SphK2 with ABC294640 inhibited TNBC lung metastasis in vivo . p21-activated kinase 1 (PAK1), p-Lin-11/Isl-1/Mec-3 kinase 1 (LIMK1) and Cofilin1 was the downstream signaling cascade of SphK2/S1P. Inhibition of PAK1 suppressed SphK2/S1P induced TNBC cells migration. Concusion: SphK2/S1P promotes TNBC metastasis through the activation of the PAK1/LIMK1/Cofilin1 signaling pathway. ABC294640 potently inhibits TNBC metastasis in vivo which could be developed as a novel agent for the clinical treatment of TNBC.

scholarly journals The Valuable Role of Holo-lactoferrin in Ferroptosis and Its Application in Radiotherapy of Triple Negative Breast Cancer (P05-014-19)

2019 ◽  
Vol 3 (Supplement_1) ◽  
Author(s):  
Zheng Zhang ◽  
Jiaying Xu ◽  
Li Qiang Qin ◽  
Yang Kai
Keyword(s):  
Breast Cancer ◽  
Cell Viability ◽  
Triple Negative Breast Cancer ◽  
Natural Science ◽  
Triple Negative ◽  
Academic Program ◽  
Jiangsu Province ◽  
Ros Generation ◽  
Mcf 7

Abstract Objectives Holo-lactoferrin (Holo-Lf) is one kind of iron-saturated form of lactoferrin. Our study is to observe the role of Holo-Lf in ferroptosis and radiotherapy of breast cancer. Methods Triple negative breast cancer (TMBC) cells MDA-MB-231 and non-TMBC cells MCF-7 cells were treated with Apo-Lf and Holo-Lf, separately. After 24 h, cell viability was determined by CCK-8. To observe the role of Holo-Lf in ferroptosis, MDA-MB-231 and MCF-7 cells were treated with Apo-Lf, Holo-Lf in combine with ferroptosis activator Erastin. After 24 h, cells were incubated with PI and ROS probe. The fluorescence intensity of PI and ROS were examined by CLSM. Besides, cells also collected for the detection of GPX-4 expression. To demonstrate the H2O2 catalytic effect of Holo-Lf, MDA-MB-231 cells were incubated with 1 mM H2O2 after treated with Holo-Lf, and followed by a 4 Gy radiation. Finally, the ROS of cells and cell viability were detected by CCK-8 and CLSM. Results Cell viability of MDA-MB-231was significantly inhibited by Holo-Lf. After combined with Erastin, the cell viability was further inhibited. Notably, the fluorescence of PI and ROS in cells treated with Holo-Lf and Erastin were higher than other groups. By contrast, Apo-Lf significant lower the fluorescence of PI in MDA-MB-231 cells and decrease the generation of ROS in MDA-MB-231 cells. However, these results were not obtained in MCF-7 cells. Additionally, the expression of GPX-4 both in MDA-MB-231 and MCF-7 cells were significant down regulated by the treatment of Holo-Lf. As a contrast, Apo-Lf could significant up regulated the expression of GPX-4 in both cells. Besides, the ROS generation of MDA-MB-231 cells treated with Holo-Lf and H2O2 combined with 4 Gy radiation were significantly higher than cells in other groups. Conclusions Holo-Lf can specially enhance the ferroptosis of Erastin to MDA-MB-231 cells. Holo-Lf also can relieve the hypoxia microenvironment and enhance the inhibition effect of radiotherapy to MDA-MB-231 cells. Funding Sources This work was supported in part by the Outstanding Youth Science Foundation, the National Natural Science Foundation of China, the National Natural Science Foundation of Jiangsu Province, Postgraduate Research & Practice Innovation Program of Jiangsu Province, and a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD). Supporting Tables, Images and/or Graphs

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