CRISPR/Cas9-mediated Bag-1 Knockout Increased Mesenchymal Characteristics of MCF-7 Cells Via Akt Hyperactivation-mediated Actin Cytoskeleton Remodeling

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2022 Jan 7

PMID 34995286

Citations 3

Authors

Pelin Ozfiliz Kilbas

Nisan Denizce Can

Tugba Kizilboga

Fikret Ezberci

Hamdi Levent Doganay

Elif Damla Arisan

Gizem Dinler Doganay

Affiliations

Soon will be listed here.

Abstract

Bag-1 protein is a crucial target in cancer to increase the survival and proliferation of cells. The Bag-1 expression is significantly upregulated in primary and metastatic cancer patients compared to normal breast tissue. Overexpression of Bag-1 decreases the efficiency of conventional chemotherapeutic drugs, whereas Bag-1 silencing enhances the apoptotic efficiency of therapeutics, mostly in hormone-positive breast cancer subtypes. In this study, we generated stable Bag-1 knockout (KO) MCF-7 breast cancer cells to monitor stress-mediated cellular alterations in comparison to wild type (wt) and Bag-1 overexpressing (Bag-1 OE) MCF-7 cells. Validation and characterization studies of Bag-1 KO cells showed different cellular morphology with hyperactive Akt signaling, which caused stress-mediated actin reorganization, focal adhesion decrease and led to mesenchymal characteristics in MCF-7 cells. A potent Akt inhibitor, MK-2206, suppressed mesenchymal transition in Bag-1 KO cells. Similar results were obtained following the recovery of Bag-1 isoforms (Bag-1S, M, or L) in Bag-1 KO cells. The findings of this study emphasized that Bag-1 is a mediator of actin-mediated cytoskeleton organization through regulating Akt activation.

Citing Articles

A BAG-1-inhibitory peptide, GO-Pep, suppresses c-Raf activity in cancer.

Tatli O, Cebi E, Turk M, Dingiloglu B, Sezan A, Basturk E Commun Biol. 2025; 8(1):336.

PMID: 40021821 PMC: 11871328. DOI: 10.1038/s42003-024-07419-4.

Bag-1-mediated HSF1 phosphorylation regulates expression of heat shock proteins in breast cancer cells.

Kizilboga T, Ozden C, Can N, Onay Ucar E, Dinler Doganay G FEBS Open Bio. 2024; 14(9):1559-1569.

PMID: 39049197 PMC: 11492399. DOI: 10.1002/2211-5463.13843.

Thio-2 Inhibits Key Signaling Pathways Required for the Development and Progression of Castration-resistant Prostate Cancer.

Neeb A, Figueiredo I, Bogdan D, Cato L, Stober J, Jimenez-Vacas J Mol Cancer Ther. 2024; 23(6):791-808.

PMID: 38412481 PMC: 11148553. DOI: 10.1158/1535-7163.MCT-23-0354.

Identification of an Imidazopyridine-based Compound as an Oral Selective Estrogen Receptor Degrader for Breast Cancer Therapy.

Pan M, Solozobova V, Kuznik N, Jung N, Grassle S, Gourain V Cancer Res Commun. 2023; 3(7):1378-1396.

PMID: 37520743 PMC: 10373600. DOI: 10.1158/2767-9764.CRC-23-0111.

References

Liu H, Wang Z, Bai Y, Wang M, Li Y, Wei S . Different BAG-1 isoforms have distinct functions in modulating chemotherapeutic-induced apoptosis in breast cancer cells. Acta Pharmacol Sin. 2009; 30(2):235-41. PMC: 4002464. DOI: 10.1038/aps.2008.21. View

Tian X, Liu Z, Niu B, Zhang J, Tan T, Lee S . E-cadherin/β-catenin complex and the epithelial barrier. J Biomed Biotechnol. 2011; 2011:567305. PMC: 3191826. DOI: 10.1155/2011/567305. View

Vandermoere F, El Yazidi-Belkoura I, Demont Y, Slomianny C, Antol J, Lemoine J . Proteomics exploration reveals that actin is a signaling target of the kinase Akt. Mol Cell Proteomics. 2006; 6(1):114-24. DOI: 10.1074/mcp.M600335-MCP200. View

Townsend P, Stephanou A, Packham G, Latchman D . BAG-1: a multi-functional pro-survival molecule. Int J Biochem Cell Biol. 2004; 37(2):251-9. DOI: 10.1016/j.biocel.2004.03.016. View

Ozfiliz P, Kizilboga T, Demir S, Alkurt G, Palavan-Unsal N, Arisan E . Bag-1 promotes cell survival through c-Myc-mediated ODC upregulation that is not preferred under apoptotic stimuli in MCF-7 cells. Cell Biochem Funct. 2015; 33(5):293-307. DOI: 10.1002/cbf.3114. View

Wahl 3rd J, Kim Y, Cullen J, Johnson K, Wheelock M . N-cadherin-catenin complexes form prior to cleavage of the proregion and transport to the plasma membrane. J Biol Chem. 2003; 278(19):17269-76. DOI: 10.1074/jbc.M211452200. View

Zhao Y, Hu X, Liu Y, Dong S, Wen Z, He W . ROS signaling under metabolic stress: cross-talk between AMPK and AKT pathway. Mol Cancer. 2017; 16(1):79. PMC: 5390360. DOI: 10.1186/s12943-017-0648-1. View

Itoh M, Nagafuchi A, Moroi S, Tsukita S . Involvement of ZO-1 in cadherin-based cell adhesion through its direct binding to alpha catenin and actin filaments. J Cell Biol. 1997; 138(1):181-92. PMC: 2139940. DOI: 10.1083/jcb.138.1.181. View

Lamouille S, Xu J, Derynck R . Molecular mechanisms of epithelial-mesenchymal transition. Nat Rev Mol Cell Biol. 2014; 15(3):178-96. PMC: 4240281. DOI: 10.1038/nrm3758. View

10.

Sousa-Squiavinato A, Vasconcelos R, Gehren A, Fernandes P, Oliveira I, Boroni M . Cofilin-1, LIMK1 and SSH1 are differentially expressed in locally advanced colorectal cancer and according to consensus molecular subtypes. Cancer Cell Int. 2021; 21(1):69. PMC: 7821653. DOI: 10.1186/s12935-021-01770-w. View

11.

Miller E, Lee H, Lulla A, Hernandez L, Gokare P, Lim B . Current treatment of early breast cancer: adjuvant and neoadjuvant therapy. F1000Res. 2014; 3:198. PMC: 4224200. DOI: 10.12688/f1000research.4340.1. View

12.

Gotz R, Wiese S, Takayama S, Camarero G, Rossoll W, Schweizer U . Bag1 is essential for differentiation and survival of hematopoietic and neuronal cells. Nat Neurosci. 2005; 8(9):1169-78. PMC: 1361767. DOI: 10.1038/nn1524. View

13.

Turk M, Tatli O, Alkan H, Ozfiliz Kilbas P, Alkurt G, Dinler Doganay G . Co-Chaperone Bag-1 Plays a Role in the Autophagy-Dependent Cell Survival through Beclin 1 Interaction. Molecules. 2021; 26(4). PMC: 7914623. DOI: 10.3390/molecules26040854. View

14.

Townsend P, Cutress R, Sharp A, Brimmell M, Packham G . BAG-1 prevents stress-induced long-term growth inhibition in breast cancer cells via a chaperone-dependent pathway. Cancer Res. 2003; 63(14):4150-7. View

15.

Kalluri R . EMT: when epithelial cells decide to become mesenchymal-like cells. J Clin Invest. 2009; 119(6):1417-9. PMC: 2689122. DOI: 10.1172/JCI39675. View

16.

Xue G, Hemmings B . PKB/Akt-dependent regulation of cell motility. J Natl Cancer Inst. 2013; 105(6):393-404. DOI: 10.1093/jnci/djs648. View

17.

Zhang X, Wang L, Zhang H, Tu F, Qiang Y, Nie C . Decreased expression of ZO-1 is associated with tumor metastases in liver cancer. Oncol Lett. 2019; 17(2):1859-1864. PMC: 6341828. DOI: 10.3892/ol.2018.9765. View

18.

Elam W, Kang H, De La Cruz E . Competitive displacement of cofilin can promote actin filament severing. Biochem Biophys Res Commun. 2013; 438(4):728-31. PMC: 3785092. DOI: 10.1016/j.bbrc.2013.07.109. View

19.

Greenwood J, Theibert A, Prestwich G, Murphy-Ullrich J . Restructuring of focal adhesion plaques by PI 3-kinase. Regulation by PtdIns (3,4,5)-p(3) binding to alpha-actinin. J Cell Biol. 2000; 150(3):627-42. PMC: 2175186. DOI: 10.1083/jcb.150.3.627. View

20.

Kalluri R, Weinberg R . The basics of epithelial-mesenchymal transition. J Clin Invest. 2009; 119(6):1420-8. PMC: 2689101. DOI: 10.1172/JCI39104. View