Positive regulation of migration and invasion by vasodilator-stimulated phosphoprotein via Rac1 pathway in human breast cancer cells
TL;DR: The data showed that the higher expression level of VASP contributed to a higher invasive migration capacity of human breast cancer cells which was controlled by the Rac1 pathway.
Abstract: This study aimed to investigate the role of the cytoskeleton-associated protein vasodilator-stimulated phosphoprotein (VASP) on the migration and invasion of human breast cancer cells and its relationship to Rac1 which is a member of the Rho family and has been found to be implicated in tumorigenesis, invasion and metastasis. We detected the mRNA and protein expression levels of VASP and Rac1 of the non-invasive breast cancer cell line MCF-7 as well as the invasive cell line MDA-MB-231 by RT-PCR and Western blotting. GST pull-down assay was used to examine the activity of Rac1. Accordingly, the cell invasive migration ability was analyzed in a wound-healing assay (2D) and transwell assays (3D migration and invasion). We then used VASP-siRNA to inhibit the expression of VASP in breast cancer cells in order to study the relationship between the VASP expression level and the invasive migration ability of breast cancer cells. Rac1-siRNA was used to decrease the expression of Rac1, and observe its effect on the VASP expression level together with the migration and invasion ability of MCF-7 and MDA-MB-231 cells. Our results revealed that the invasive breast cancer cell line MDA-MB-231 showed a higher Rac1 activity and VASP expression level compared with the non-invasive MCF-7. Inhibition of Rac1 or VASP by siRNA, respectively, decreased the migration and invasion ability of breast cancer cells and the transfection of Rac1 siRNA-mediated reduction of VASP expression at mRNA and protein levels. Collectively, our data showed that the higher expression level of VASP contributed to a higher invasive migration capacity of human breast cancer cells which was controlled by the Rac1 pathway.
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TL;DR: The data reveal a critical role of VASP in non‐tumor cells in the tumor environment for melanoma growth in vivo, and Histological analyses showed smaller cells with impaired nutrition status and less vascularization in melanomas derived from VASp−/− versus counterparts from wild‐type mice.
8 citations
Cites background from "Positive regulation of migration an..."
...[9,10] and VASP expression is up-regulated in human lung carcinoma and increases with more advanced tumor stages....
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...Consistently, elevated VASP expression levels increase invasive migration of human breast cancer cells [10]....
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...Inhibition of VASP by siRNA showed that the higher expression levels of VASP contribute to higher invasive migration capacity of the human breast cancer cell line MDA-MB-231 [10]....
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27 Feb 2013
TL;DR: The anti-apoptotic, pro-angiogenic effects of PI3K/Akt/mTOR may be mediated, at least in part, through a downstream signaling pathway involving en‐ dogenous endothelial-form nitric oxide synthase (eNOS, also called NOS3), and subsequent‐ ly soluble guanylyl cyclase (sGC) and protein kinase G (PKG).
Abstract: Platinum-based drugs such as cisplatin (cis-diammine-dichloro-platinum, also commonly known as CDDP) have dominated the drug therapy of ovarian cancer during the past three decades [1]. Cisplatin interacts with DNA to form intrastrand crosslink adducts, and its mo‐ lecular mechanism involves regulation of p53 and the mitogen-activated protein kinase (MAPK) signaling pathway [2]. The phosphatidylinositol-3-kinase (PI3K)/Akt signaling pathway is crucial for regulation of survival and for progression and chemoresistance in ovarian cancer, leading to the development of new chemotherapeutic inhibitors targeting the PI3K/Akt pathway and the downstream serine/threonine protein kinase mTOR. [3]. In‐ hibition of PI3K pathway signaling using PI3K or mTOR inhibitors has been shown to sensi‐ tize ovarian cancer cell lines to the apoptosis-inducing effect of platinum compounds [4, 5]. In addition, activation of the PI3K/Akt/mTOR pathway in ovarian cancer cell lines contrib‐ utes to cisplatin resistance [6]. The anti-apoptotic, pro-angiogenic effects of PI3K/Akt/mTOR may be mediated, at least in part, through a downstream signaling pathway involving en‐ dogenous endothelial-form nitric oxide synthase (eNOS, also called NOS3), and subsequent‐ ly soluble guanylyl cyclase (sGC) and protein kinase G (PKG). Studies have shown that Akt activates eNOS by phosphorylating human eNOS at Ser1177 (equivalent to bovine eNOS at Ser1179), leading to an increase in nitric oxide (NO) production in endothelial cells [7, 8]. In the cases of vascular endothelial growth factor (VEGF) [9, 10], sphingosine 1-phosphate [11, 12], and estrogen [13, 14], there are vast evidences suggesting PI3K-activation of Akt is re‐ sponsible for regulating the phosphorylation and activation of eNOS. In bovine aortic endo‐ thelial cells, eNOS co-immunoprecipitates with Akt, indicating that the two enzymes associate in vivo, and Akt directly activates eNOS, increasing eNOS activity by 15-20 fold
6 citations
01 Apr 2002
TL;DR: In this article, it was shown that Ena/VASP proteins are highly expressed in the developing cortical plate in cells bordering Reelin-expressing Cajal-Retzius cells and in the intermediate zone through which newly born cells migrate.
Abstract: Development of the multilayered cerebral cortex involves extensive regulated migration of neurons arising from the deeper germinative layers of the mammalian brain [1]. The anatomy and formation of the cortical layers has been well characterized; however, the underlying molecular mechanisms that control the migration and the final positioning of neurons within the cortex remain poorly understood [2, 3]. Here, we report evidence for a key role of Ena/VASP proteins, a protein family implicated in the spatial control of actin assembly [4] and previously shown to negatively regulate fibroblast cell speeds [5], in cortical development. Ena/VASP proteins are highly expressed in the developing cortical plate in cells bordering Reelin-expressing Cajal-Retzius cells and in the intermediate zone through which newly born cells migrate. Inhibition of Ena/VASP function through retroviral injections in utero led to aberrant placement of early-born pyramidal neurons in the superficial layers of both the embryonic and the postnatal cortex in a cell-autonomous fashion. The abnormally placed pyramidal neurons exhibited grossly normal morphology and polarity. Our results are consistent with a model in which Ena/VASP proteins function in vivo to control the position of neurons in the mouse neocortex.
5 citations
TL;DR: It is concluded from this study that the present strategy is capable of selecting perturbed molecular functions that putatively play roles in the progression of diseases and provides an improved interpretability of GO terms based on the definition of Gene Ontology codes.
Abstract: A systems biology approach for the identification of perturbed molecular functions is required to understand the complex progressive disease such as breast cancer. In this study, we analyze the microarray data with Gene Ontology terms of molecular functions to select perturbed molecular functional modules in breast cancer tissues based on the definition of Gene ontology Functional Code. The Gene Ontology is three structured vocabularies describing genes and its products in terms of their associated biological processes, cellular components and molecular functions. The Gene Ontology is hierarchically classified as a directed acyclic graph. However, it is difficult to visualize Gene Ontology as a directed tree since a Gene Ontology term may have more than one parent by providing multiple paths from the root. Therefore, we applied the definition of Gene Ontology codes by defining one or more GO code(s) to each GO term to visualize the hierarchical classification of GO terms as a network. The selected molecular functions could be considered as perturbed molecular functional modules that putatively contributes to the progression of disease. We evaluated the method by analyzing microarray dataset of breast cancer tissues; i.e., normal and invasive breast cancer tissues. Based on the integration approach, we selected several interesting perturbed molecular functions that are implicated in the progression of breast cancers. Moreover, these selected molecular functions include several known breast cancer-related genes. It is concluded from this study that the present strategy is capable of selecting perturbed molecular functions that putatively play roles in the progression of diseases and provides an improved interpretability of GO terms based on the definition of Gene Ontology codes.
1 citations
Additional excerpts
...…(Kulka et al., 2009), F3/TF (Amirkhosravi et al., 1998), FZD1 (Benhaj et al., 2006), NGFR (Reis-Filho et al., 2006), LEFR (Han et al., 2008), RAC1 (Han et al., 2008), PLXNB1 (Rody et al., 2007), IGHG1 (Kabbage et al., 2008), IGHG3 (Bin Amer et al., 2008), TLR3 (Salaun et al., 2006), FBN1 (Chen et…...
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...…et al., 2007), CLDN4 (Kulka et al., 2009), F3/TF (Amirkhosravi et al., 1998), FZD1 (Benhaj et al., 2006), NGFR (Reis-Filho et al., 2006), LEFR (Han et al., 2008), RAC1 (Han et al., 2008), PLXNB1 (Rody et al., 2007), IGHG1 (Kabbage et al., 2008), IGHG3 (Bin Amer et al., 2008), TLR3 (Salaun et…...
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...Moreover, there are considerable amount of genes in Table 5 that are implicated to play roles in the progression of breast cancer or other cancers, including BDKRB2 (Greco et al., 2005), PLCB1 (Naor, 2009), PLD1 (Eisen and Brown, 2002), GPLD1 (Derevianko et al., 1996; Williams et al., 2001), EXO1 (Naderi et al., 2007), ANG (Campo et al., 2005), ISG20/HEM45 (Pentecost, 1998), AZGP1/ZAG (Hassan et al., 2009), ZKSCAN1 (Pennanen et al., 2009), REXO2 (Flanagan et al., 2009), CTDSPL (Murabito et al., 2007), NT5E (Zhou et al., 2007), DLG1 (Fuja et al., 2004), PPM1A (Lin et al., 2006), TIAM1 (Lane et al., 2008), BAG4 (Yang et al., 2006), BRAF (Hollestelle, et al., 2007), CLDN4 (Kulka et al., 2009), F3/TF (Amirkhosravi et al., 1998), FZD1 (Benhaj et al., 2006), NGFR (Reis-Filho et al., 2006), LEFR (Han et al., 2008), RAC1 (Han et al., 2008), PLXNB1 (Rody et al., 2007), IGHG1 (Kabbage et al., 2008), IGHG3 (Bin Amer et al., 2008), TLR3 (Salaun et al., 2006), FBN1 (Chen et al., 2007), DUSP4 (Venter et al., 2005), PTPN3 (Wang et al., 2004), CDKN3/KAP (Lee et al., 2000), and etc. Based on the integration of GFC codes and microarray data of breast cancer tissues, we present several perturbed molecular functions, which are implicated in the progression of breast cancer....
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..., 2008), RAC1 (Han et al., 2008), PLXNB1 (Rody et al....
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Patent•
07 Aug 2014
TL;DR: In this article, the state of an epithelial tumor or tissue in a subject is detected by measuring the amount of pSer239-VASP, pSerl57-VSP, and/or total VASP in the subject or in a tissue or tumor of a subject.
Abstract: Disclosed herein are methods for diagnosing the state of an epithelial tumor or tissue in a subject. Also disclosed are methods of diagnosing tumor initiation in an epithelial tissue of a subject. Also disclosed are methods of determining prognosis of a subject with an epithelial tumor. These methods include, in part, measuring the amount of pSer239-VASP, pSerl57-VASP, and/or total VASP in a subject or in a tissue or tumor of a subject and comparing those levels to an appropriate reference level. This abstract is intended as a scanning tool for purposes of searching in the particular art and is not intended to be limiting of the present invention.
1 citations
References
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TL;DR: 21-nucleotide siRNA duplexes provide a new tool for studying gene function in mammalian cells and may eventually be used as gene-specific therapeutics.
Abstract: RNA interference (RNAi) is the process of sequence-specific, post-transcriptional gene silencing in animals and plants, initiated by double-stranded RNA (dsRNA) that is homologous in sequence to the silenced gene. The mediators of sequence-specific messenger RNA degradation are 21- and 22-nucleotide small interfering RNAs (siRNAs) generated by ribonuclease III cleavage from longer dsRNAs. Here we show that 21-nucleotide siRNA duplexes specifically suppress expression of endogenous and heterologous genes in different mammalian cell lines, including human embryonic kidney (293) and HeLa cells. Therefore, 21-nucleotide siRNA duplexes provide a new tool for studying gene function in mammalian cells and may eventually be used as gene-specific therapeutics.
10,451 citations
"Positive regulation of migration an..." refers background in this paper
...It has been found that 21 short nucleotide (nt) ds RNA molecules, known as short interfering RNAs (siRNAs), can mediate RNAi in mammalian cell lines (19)....
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TL;DR: This work challenges previous assumptions about how the G1/S transition of the mammalian cell cycle is governed, helps explain some enigmatic features of cell cycle control that also involve the functions of the retinoblastoma protein (Rb) and the INK4 proteins, and changes the thinking about how either p16 loss or overexpression of cyclin D-dependent kinases contribute to cancer.
Abstract: Mitogen-dependent progression through the first gap phase (G1) and initiation of DNA synthesis (S phase) during the mammalian cell division cycle are cooperatively regulated by several classes of cyclin-dependent kinases (CDKs) whose activities are in turn constrained by CDK inhibitors (CKIs). CKIs that govern these events have been assigned to one of two families based on their structures and CDK targets. The first class includes the INK4 proteins (inhibitors of CDK4), so named for their ability to specifically inhibit the catalytic subunits of CDK4 and CDK6. Four such proteins [p16 (Serrano et al. 1993), p15 (Hannon and Beach 1994), p18 (Guan et al. 1994; Hirai et al. 1995), and p19 (Chan et al. 1995; Hirai et al. 1995)] are composed of multiple ankyrin repeats and bind only to CDK4 and CDK6 but not to other CDKs or to D-type cyclins. The INK4 proteins can be contrasted with more broadly acting inhibitors of the Cip/Kip family whose actions affect the activities of cyclin D-, E-, and A-dependent kinases. The latter class includes p21 (Gu et al. 1993; Harper et al. 1993; El-Deiry et al. 1993; Xiong et al. 1993a; Dulic et al. 1994; Noda et al. 1994), p27 (Polyak et al. 1994a,b; Toyoshima and Hunter 1994), and p57 (Lee et al. 1995; Matsuoka et al. 1995), all of which contain characteristic motifs within their amino-terminal moieties that enable them to bind both to cyclin and CDK subunits (Chen et al. 1995, 1996; Nakanishi et al. 1995; Warbrick et al. 1995; Lin et al. 1996; Russo et al. 1996). Based largely on in vitro experiments and in vivo overexpression studies, CKIs of the Cip/Kip family were initially thought to interfere with the activities of cyclin D-, E-, and A-dependent kinases. More recent work has altered this view and revealed that although the Cip/Kip proteins are potent inhibitors of cyclin Eand A-dependent CDK2, they act as positive regulators of cyclin Ddependent kinases. This challenges previous assumptions about how the G1/S transition of the mammalian cell cycle is governed, helps explain some enigmatic features of cell cycle control that also involve the functions of the retinoblastoma protein (Rb) and the INK4 proteins, and changes our thinking about how either p16 loss or overexpression of cyclin D-dependent kinases contribute to cancer. Here we focus on the biochemical interactions that occur between CKIs and cyclin Dand E-dependent kinases in cultured mammalian cells, emphasizing the manner by which different positive and negative regulators of the cell division cycle cooperate to govern the G1-to-S transition. To gain a more comprehensive understanding of the biology of CDK inhibitors, readers are encouraged to refer to a rapidly emerging but already extensive literature (for review, see Elledge and Harper 1994; Sherr and Roberts 1995; Chellappan et al. 1998; Hengst and Reed 1998a; Kiyokawa and Koff 1998; Nakayama 1998; Ruas and Peters 1998).
6,076 citations
"Positive regulation of migration an..." refers background in this paper
...As a significant member of the Rho family, Rac1 has been implicated in tumorigenesis (2), tumor angiogenesis (3), invasion and metastasis (4), cell-cycle control and apoptosis (5)....
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TL;DR: In this review, functions of small G proteins and their modes of activation and action are described.
Abstract: Small GTP-binding proteins (G proteins) exist in eukaryotes from yeast to human and constitute a superfamily consisting of more than 100 members. This superfamily is structurally classified into at least five families: the Ras, Rho, Rab, Sar1/Arf, and Ran families. They regulate a wide variety of cell functions as biological timers (biotimers) that initiate and terminate specific cell functions and determine the periods of time for the continuation of the specific cell functions. They furthermore play key roles in not only temporal but also spatial determination of specific cell functions. The Ras family regulates gene expression, the Rho family regulates cytoskeletal reorganization and gene expression, the Rab and Sar1/Arf families regulate vesicle trafficking, and the Ran family regulates nucleocytoplasmic transport and microtubule organization. Many upstream regulators and downstream effectors of small G proteins have been isolated, and their modes of activation and action have gradually been elucidated. Cascades and cross-talks of small G proteins have also been clarified. In this review, functions of small G proteins and their modes of activation and action are described.
2,520 citations
Additional excerpts
...progression and cell-cell adhesion (1)....
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TL;DR: Pure components of the actin cytoskeleton are used to reconstitute sustained movement in Listeria and Shigella in vitro and have implications for the understanding of the mechanism of actin-based motility in cells.
Abstract: Actin polymerization is essential for cell locomotion and is thought to generate the force responsible for cellular protrusions. The Arp2/3 complex is required to stimulate actin assembly at the leading edge in response to signalling. The bacteria Listeria and Shigella bypass the signalling pathway and harness the Arp2/3 complex to induce actin assembly and to propel themselves in living cells. However, the Arp2/3 complex alone is insufficient to promote movement. Here we have used pure components of the actin cytoskeleton to reconstitute sustained movement in Listeria and Shigella in vitro. Actin-based propulsion is driven by the free energy released by ATP hydrolysis linked to actin polymerization, and does not require myosin. In addition to actin and activated Arp2/3 complex, actin depolymerizing factor (ADF, or cofilin) and capping protein are also required for motility as they maintain a high steady-state level of G-actin, which controls the rate of unidirectional growth of actin filaments at the surface of the bacterium. The movement is more effective when profilin, alpha-actinin and VASP (for Listeria) are also included. These results have implications for our understanding of the mechanism of actin-based motility in cells.
959 citations
"Positive regulation of migration an..." refers background in this paper
...Furthermore, the addition of VASP resulted in an increase in bacterial speed in the in vitro reconstitution assays of Listeria motility (34)....
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...That VASP can affect the cell movements of bacteria, mouse melanoma cells, human platelet, fibroblasts and neurons by regulating cytoskeleton has been detected (14,15,34,35)....
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TL;DR: A spatial inventory of the many molecular players in this dynamic domain of the actin cytoskeleton is given in order to highlight the open questions and the challenges ahead.
939 citations
"Positive regulation of migration an..." refers background in this paper
...During cell migration, activated Rac and Cdc42 induce reorganization of the actin cytoskeleton at the leading edge (6)....
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