Mutant p53 Disrupts Role of ShcA Protein in Balancing Smad Protein-dependent and -independent Signaling Activity of Transforming Growth Factor-β (TGF-β)

Lin, Shu Chun; Yu, Lan; Yang, Junhua; Zhao, Lijun; Karia, Bijal; Bishop, Alexander J.R.; Jackson, James G.; Lozano, Guillermina; Copland, John A.; Mu, Xiaoxin; Sun, Beicheng; Sun, LuZhe

doi:10.1074/jbc.m111.265397

Cited by 13 publications

(16 citation statements)

References 53 publications

(39 reference statements)

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“…Treatment with combination TGFβ-dasatinib did not have a significant effect on the non-canonical TGFβ pathway intracellular mediators ERK, AKT, or p38 ( S3 Figure ). In addition, treatment with TGFβ-1 did not result in an increase in Shc activation in A549 cells, as previously reported for other cell types [42] , [43] , and knockdown of the expression of ShcA using ShcA siRNA did not have an effect in apoptosis in ShcA-negative A549 cells (data not shown), suggesting that this pathway is not involved in the induced apoptosis observed with TGFβ-dasatinib treatment.…”

Section: Resultssupporting

confidence: 86%

Transforming Growth Factor β Signaling Overcomes Dasatinib Resistance in Lung Cancer

Gordián

Pevzner

et al. 2014

PLoS ONE

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Lung cancer is the second most common cancer and the leading cause of cancer-related deaths. Despite recent advances in the development of targeted therapies, patients with advanced disease remain incurable, mostly because metastatic non-small cell lung carcinomas (NSCLC) eventually become resistant to tyrosine kinase inhibitors (TKIs). Kinase inhibitors have the potential for target promiscuity because the kinase super family is the largest family of druggable genes that binds to a common substrate (ATP). As a result, TKIs often developed for a specific purpose have been found to act on other targets. Drug affinity chromatography has been used to show that dasatinib interacts with the TGFβ type I receptor (TβR-I), a serine-threonine kinase. To determine the potential biological relevance of this association, we studied the combined effects of dasatinib and TGFβ on lung cancer cell lines. We found that dasatinib treatment alone had very little effect; however, when NSCLC cell lines were treated with a combination of TGFβ and dasatinib, apoptosis was induced. Combined TGFβ-1 + dasatinib treatment had no effect on the activity of Smad2 or other non-canonical TGFβ intracellular mediators. Interestingly, combined TGFβ and dasatinib treatment resulted in a transient increase in p-Smad3 (seen after 3 hours). In addition, when NSCLC cells were treated with this combination, the pro-apoptotic protein BIM was up-regulated. Knockdown of the expression of Smad3 using Smad3 siRNA also resulted in a decrease in BIM protein, suggesting that TGFβ-1 + dasatinib-induced apoptosis is mediated by Smad3 regulation of BIM. Dasatinib is only effective in killing EGFR mutant cells, which is shown in only 10% of NSCLCs. Therefore, the observation that wild-type EGFR lung cancers can be manipulated to render them sensitive to killing by dasatinib could have important implications for devising innovative and potentially more efficacious treatment strategies for this disease.

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Section: Resultssupporting

confidence: 86%

Transforming Growth Factor β Signaling Overcomes Dasatinib Resistance in Lung Cancer

Gordián

Pevzner

et al. 2014

PLoS ONE

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“…Cells were seeded in a 96-well plate at 2,000 cells/well with or without TGF-β3 (0.5 ng/ml) treatment at various time points and MTT assay was performed as described previously [36]. Two hours before each time point, 3-19(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) (2mg/ml in PBS) was added at 50 μl per well and cells were incubated at 37°C for 2 hours.…”

Section: Methodsmentioning

confidence: 99%

Blockade of Autocrine TGF-β Signaling Inhibits Stem Cell Phenotype, Survival, and Metastasis of Murine Breast Cancer Cells

Liu¹,

Bandyopadhyay

Nichols³

et al. 2012

J Stem Cell Res Ther

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Transforming growth factor beta (TGF-β) signaling has been implicated in driving tumor progression and metastasis by inducing stem cell-like features in some human cancer cell lines. In this study, we have utilized a novel murine cell line NMuMG-ST, which acquired cancer stem cell (CSC) phenotypes during spontaneous transformation of the untransformed murine mammary cell line NMuMG, to investigate the role of autocrine TGF-β signaling in regulating their survival, metastatic ability, and the maintenance of cancer stem cell characteristics. We have retrovirally transduced a dominant-negative TGF-β type II receptor (DNRII) into the NMuMG-ST cell to abrogate autocrine TGF-β signaling. The expression of DNRII reduced TGF-β sensitivity of the NMuMG-ST cells in various cell-based assays. The blockade of autocrine TGF-β signaling reduced the ability of the cell to grow anchorage-independently and to resist serum deprivation-induced apoptosis. These phenotypes were associated with reduced levels of active and phosphorylated AKT and ERK, and Gli1 expression suggesting that these pathways contribute to the growth and survival of this model system. More interestingly, the abrogation of autocrine TGF-β signaling also led to the attenuation of several features associated with mammary stem cells including epithelial-mesenchymal transition, mammosphere formation, and expression of stem cell markers. When xenografted in athymic nude mice, the DNRII cells were also found to undergo apoptosis and induced significantly lower lung metastasis burden than the control cells even though they formed similar size of xenograft tumors. Thus, our results indicate that autocrine TGF-β signaling is involved in the maintenance and survival of stem-like cell population resulting in the enhanced metastatic ability of the murine breast cancer cells.

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“…3A). To determine the role of TβRII and consequently that of the TGF-β signaling pathway in regulating the malignant phenotypes of HCC cells, we knocked down TβRII in SNU423 and Sk-Hep-1 cells with the stable expression of a TβRII shRNA as described previously [22]. Results indicated that knockdown of TβRII, confirmed by Western blotting analysis and RT-PCR (Fig.…”

Section: Resultsmentioning

confidence: 81%

“…The sequence of shRNA is: TGFBRII: 5′-CCG GCC TGA CTT GTT GCT AGT CAT ACT CGA GTA TGA CTA GCA ACA AGT CAG GTT TTT G-3′ as described previously [22]; Smad4∶5′-CCG GCG AGT TGT ATC ACC TGG AAT TCT CGA GAA TTC CAG GTG ATA CAA CTC GTT TTT G-3′. The virus containing TβRII shRNA, Smad4 shRNA or control shRNA was produced by transfecting HEK 293FT packaging cells with shRNA expression plasmids in Lipofectamine 2000 according to the manufacture’s protocol.…”

Section: Methodsmentioning

confidence: 99%

TGF-β Signaling Is Often Attenuated during Hepatotumorigenesis, but Is Retained for the Malignancy of Hepatocellular Carcinoma Cells

Lin

Yang

et al. 2013

PLoS ONE

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The role of transforming growth factor-beta (TGF-β) signaling in hepatocarcinogenesis remains controversial. We aimed to reveal TGF-β signaling status in human and murine tissues of hepatocellular carcinoma (HCC) and the mechanisms that mediate TGF-β’s role in regulating HCC malignancy. Here, TGF-β pathway component expression and activation in human and murine HCC tissues were measured with quantitative RT-PCR and Western blotting assays. The role of TGF-β receptor and Smad signaling in the growth and survival of several HCC cell lines was determined with several in vitro and in vivo approaches. We found that TGF-β receptor II (TβRII) expression was downregulated in two different HCC patient cohorts. Consistently, Smad3 phosphorylation was also downregulated in HCC tissues in comparison to that in adjacent normal tissues. Interestingly, many HCC cell lines were sensitive to TGF-β and growth-inhibited by exogenous TGF-β. However, stable knockdown of TβRII inhibited cell growth on plastic and in soft agar, and induced apoptosis resulting in suppressed subcutaneous tumor growth and metastatic potential in vivo. Furthermore, knockdown of Smad4 also led to a significant inhibition of growth on plastic and in soft agar with concomitant increase of apoptosis, PTEN expression, and reduced nuclear accumulation of linker region-phosphorylated Smad3. Taken together, TGF-β signaling pathway plays a dichotomous role in hepatocellular carcinogenesis. It appears to suppress HCC development, but is retained for HCC cell survival and malignancy. Furthermore, Smad4 can mediate both growth inhibitory activity induced by exogenous TGF-β and the survival activity induced by autocrine TGF-β revealing a delicate selection of the two opposing activities of TGF-β during HCC evolution.

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Mutant p53 Disrupts Role of ShcA Protein in Balancing Smad Protein-dependent and -independent Signaling Activity of Transforming Growth Factor-β (TGF-β)

Cited by 13 publications

References 53 publications

Transforming Growth Factor β Signaling Overcomes Dasatinib Resistance in Lung Cancer

Transforming Growth Factor β Signaling Overcomes Dasatinib Resistance in Lung Cancer

Blockade of Autocrine TGF-β Signaling Inhibits Stem Cell Phenotype, Survival, and Metastasis of Murine Breast Cancer Cells

TGF-β Signaling Is Often Attenuated during Hepatotumorigenesis, but Is Retained for the Malignancy of Hepatocellular Carcinoma Cells

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