Axl activates autocrine transforming growth factor‐β signaling in hepatocellular carcinoma

Reichl, Patrick; Dengler, Mirko; Zijl, Franziska van; Huber, Heidemarie; Führlinger, Gerhard; Reichel, Christian; Sieghart, Wolfgang; Peck‐Radosavljevic, Markus; Grubinger, Markus; Mikulits, Wolfgang

doi:10.1002/hep.27492

Cited by 138 publications

(188 citation statements)

References 31 publications

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“…Increasing evidence has suggested that signaling pathways such as WNT/β-catenin and TGFβ/SMAD pathways may be involved in modulating tumorigenesis [33, 34]. Among these pathways, activation of TGFβ/SMAD plays an important role in enhancing the EMT to remodel the prometastasis phenotype [11], and SNAIL family members are regulated by the TGFβ/SMAD signaling pathway [22].…”

Section: Resultsmentioning

confidence: 99%

MEGF6 Promotes the Epithelial-to-Mesenchymal Transition via the TGFβ/SMAD Signaling Pathway in Colorectal Cancer Metastasis

Hu¹,

Wang²,

Wang³

et al. 2018

Cell Physiol Biochem

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Background/Aims: Colorectal cancer (CRC) is a malignancy that has high morbidity and mortality and is initiated from accumulative genetic events. Although much effort has been made to elucidate the genetic mechanism underlying this disease, it still remains unknown. Here, we discovered a novel role for multiple epidermal growth factor-like domains protein 6 (MEGF6) in CRC, namely, that it induces the epithelial-to-mesenchymal transition (EMT) to promote CRC metastasis via the transforming growth factor beta (TGFβ)/SMAD signaling pathway. Methods: RNA sequencing data from the Gene Expression Omnibus database were analyzed using R software. Based on The Cancer Genome Atlas Colon Adenocarcinoma (TCGA-COAD) cohort, the clinical significance of MEGF6 was investigated. HCT8R, HCT116, and LoVo CRC cells were transfected with small interfering RNA against MEGF6, and their proliferation and sensitivity to fluorouracil were evaluated with the MTT cell proliferation and colony formation assays. Proteins associated with cell growth were detected by western blot analysis. The apoptosis of cells was evaluated by Annexin V/propidium iodide staining, and transwell assays were performed to assess the involvement of MEGF6 in cell migration. Markers of EMT and TGFβ/SMAD signaling were evaluated by quantitative PCR and western blotting, and the correlation between MEGF6 and these markers was assessed in the TCGA colon and renal adenocarcinoma cohort. Results: The results showed that MEGF6 was upregulated in HCT8R cells. In addition, MEGF6 was significantly overexpressed in tumor tissue and predicted a poor survival in the TCGA-COAD cohort. Moreover, MEGF6 accelerated CRC cell growth and inhibited apoptosis, and promoted CRC metastasis by inducing the EMT. Finally, we found that TGFβ/SMAD signaling triggered the expression of Slug, which regulates the MEGF6-mediated EMT. Conclusions: MEGF6 may serve as an oncogene to promote cell proliferation and inhibit apoptosis. MEGF6 can also accelerate cell migration via TGFβ/SMAD signaling-mediated EMT.

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Section: Resultsmentioning

confidence: 99%

MEGF6 Promotes the Epithelial-to-Mesenchymal Transition via the TGFβ/SMAD Signaling Pathway in Colorectal Cancer Metastasis

Hu¹,

Wang²,

Wang³

et al. 2018

Cell Physiol Biochem

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“…Recently, AXL upregulation and activation by GAS6 has been implicated in the EMT of breast cancer and hepatocellular carcinoma (28,29). Likewise, the expression of both AXL and GAS6 is deemed to be closely related to full-blown EMT in a subset of lung AD.…”

Section: Univariate Analysis Multivariate Analysismentioning

confidence: 99%

AXL and GAS6 co-expression in lung adenocarcinoma as a prognostic classifier

et al. 2017

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Abstract. AXL, a receptor tyrosine kinase implicated in cell survival, proliferation, and migration, is also associated with acquired resistance to epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor therapy. However, its prognostic significance in lung adenocarcinoma (AD) remains unclear. We therefore evaluated the prognostic significance of the expression of AXL and/or its ligand, growth arrestspecific 6 (GAS6), in completely resected lung AD. We evaluated the relationship between AXL, GAS6, and vimentin expression, as determined by immunohistochemistry (IHC) analysis, with overall survival and disease-free survival in 113 patients with stages I-III lung AD. Protein expression was also assayed using western blot analysis in 10 lung AD cell lines. AXL-positive (AXL

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“…Our recent data demonstrated that the siRNA-mediated loss of Axl expression caused a decrease of cell motility of human hepatoma cells (27). Therefore, we next analyzed the effects of the genomic editing events on the migratory cell behavior by performing a wound healing assay.…”

Section: Migration Of Crispr/cas9-edited Hepatoma Cellsmentioning

confidence: 99%

“…Chromosomal abnormalities occur in 75% of blood cancers and in more than 90% of solid tumors including hepatocellular carcinoma (HCC) (25,26). The overexpression of the receptor tyrosine kinase Axl along with the release of soluble Axl (sAxl) was detected in HCC and correlated with poor survival of HCC patients (27,28). In this study, we have generated knockouts of Axl in hepatoma cell lines using NHEJ-mediated CRISPR/Cas9 knockout technology.…”

Section: Introductionmentioning

confidence: 99%

Dynamics of CRISPR/Cas9-mediated genomic editing of the AXL locus in hepatocellular carcinoma cells

et al. 2017

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Abstract. Genomic editing using the CRISPR/Cas9 technology allows selective interference with gene expression. With this method, a multitude of haploid and diploid cells from different organisms have been employed to successfully generate knockouts of genes coding for proteins or small RNAs. Yet, cancer cells exhibiting an aberrant ploidy are considered to be less accessible to CRISPR/Cas9-mediated genomic editing, as amplifications of the targeted gene locus could hamper its effectiveness. Here we examined the suitability of CRISPR/Cas9 to knockout the receptor tyrosine kinase Axl in the human hepatoma cell lines HLF and SNU449. The genomic editing events were validated in two single cell clones each from putative HLF and SNU449 knockout cells (HLF-Axl --1, HLF-Axl --2, SNU449-Axl --1, SNU449-Axl --2). Sequence analysis of respective AXL loci revealed one to six editing events in each individual Axl -clone. The majority of insertions and deletions in the AXL gene at exon 7/8 resulted in a frameshift and thus a premature stop in the coding region.However, one genomic editing event led to an insertion of two amino acids resulting in an altered protein sequence rather than in a frameshift in the AXL locus of the SNU449-Axl --1 cells. Notably, while no Axl protein expression could be detected by immunoblotting in all four cell clones, both expression of total Axl as well as release of soluble Axl into the supernatant was observed by ELISA in incompletely edited SNU449-Axl --1 cells. Importantly, a comparative genomic hybridization array revealed comparable genomic changes in Axl knockout cells as well as in cells expressing Cas9 nickase without guide RNAs in SNU449 and HLF cells, indicating vast alterations in genomic DNA triggered by nickase. Together, these data show that the dynamics of CRISPR/Cas9 may cause incomplete editing events in cancer cell lines, as gene copy numbers vary based on genomic heterogeneity.

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Axl activates autocrine transforming growth factor‐β signaling in hepatocellular carcinoma

Abstract: Our data suggest that Axl/14-3-3ζ signaling is central for TGF-β-mediated HCC progression and a promising target for HCC therapy.

Cited by 138 publications

References 31 publications

MEGF6 Promotes the Epithelial-to-Mesenchymal Transition via the TGFβ/SMAD Signaling Pathway in Colorectal Cancer Metastasis

MEGF6 Promotes the Epithelial-to-Mesenchymal Transition via the TGFβ/SMAD Signaling Pathway in Colorectal Cancer Metastasis

AXL and GAS6 co-expression in lung adenocarcinoma as a prognostic classifier

Dynamics of CRISPR/Cas9-mediated genomic editing of the AXL locus in hepatocellular carcinoma cells

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