Rnd3/RhoE expression is regulated by G-actin through MKL1-SRF signaling pathway

Piquet, Léo; Robbe, Terezinha; Neaud, Véronique; Basbous, Sara; Rosciglione, Stéphanie; Saltel, Frédéric; Moreau, Violaine

doi:10.1016/j.yexcr.2018.06.023

Cited by 7 publications

(8 citation statements)

References 27 publications

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“…DNA and siRNA transfections were realized as described previously 11 . pEGFP-Fascin-1 vector was a generous gift from Dr D. Vignjevic (Paris, France).…”

Section: Transfectionmentioning

confidence: 99%

“…HB cell line Huh6 was generously provided by C. Perret (Paris, France). All cell lines were cultured as previously described 10,11 .…”

Section: Cell Culturementioning

confidence: 99%

“…Assays were performed as previously described 11 . pGL4-TOP reporter with TCF responsive elements to quantify ß-catenin transcriptional activity was a generous gift from Pr.…”

Section: Luciferase Reporter Assaysmentioning

confidence: 99%

“…Cell lysates were cleared of cellular debris and nuclei by a 16,000 × g centrifugation step for 10 min. Lysates were analyzed using a western-blot protocol described previously 11 . The antibodies used are listed in Table S1.…”

Section: Western Blotmentioning

confidence: 99%

“…RNA was collected from cultured cells using the Trizol reagent (Invitrogen), according to manufacturer's protocol. Reverse transcription and SYBR® Green-based real-time PCR were performed as described previously 11 . Gene expression results were first normalized to internal control r18S.…”

Section: Quantitative Real-time Pcrmentioning

confidence: 99%

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The ß-catenin-target Fascin-1, altering hepatocyte differentiation, is a new marker of immature cells in hepatoblastomas

Gest

Sena

Neaud

et al. 2021

Preprint

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BACKGROUND & AIMSß-catenin is a well-known effector of the Wnt pathway and a key player in cadherin-mediated cell adhesion. Oncogenic mutations of ß-catenin are highly frequent in pediatric liver primary tumors. Those mutations are mostly heterozygous allowing the co-expression of wild-type (WT) and mutated ß-catenins in tumor cells. We investigated the interplay between WT and mutated ß-catenins in liver tumor cells, and searched for new actors of the ß-catenin pathway.METHODSUsing an RNAi strategy in ß-catenin-mutated hepatoblastoma (HB) cells, we dissociated the structural and transcriptional activities of β-catenin, carried mainly by, respectively, WT and mutated proteins. Their impact was characterized using transcriptomic and functional analyses. We studied mice that develop liver tumors upon activation of ß-catenin in hepatocytes (APCKO and ß-cateninΔexon3 mice). We made use of transcriptomic data from mouse and human HB specimens and analyzed samples by immunohistochemistry.RESULTSWe highlighted an antagonist role of WT and mutated ß-catenins on hepatocyte differentiation as attested by alteration of hepatocyte markers expression and bile canaliculi formation. We characterized Fascin-1 as a target of ß-catenin involved in hepatocyte differentiation. Using mouse models that allow the formation of two phenotypically distinct tumors (differentiated or undifferentiated), we found that Fascin-1 expression is higher in undifferentiated tumors. Finally, we found that Fascin-1 is a specific marker of the embryonal component in human HBs.CONCLUSIONSIn mice and human, Fascin-1 expression is linked to loss of differentiation and polarity of hepatocytes. Thus, we highlighted Fascin-1 as a new player in the modulation of hepatocyte differentiation associated to ß-catenin pathway alteration in the liver.Data Transparency Statementstudy materials will be made available to other researchers upon request.

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“…DNA and siRNA transfections were realized as described previously 11 . pEGFP-Fascin-1 vector was a generous gift from Dr D. Vignjevic (Paris, France).…”

Section: Transfectionmentioning

confidence: 99%

“…HB cell line Huh6 was generously provided by C. Perret (Paris, France). All cell lines were cultured as previously described 10,11 .…”

Section: Cell Culturementioning

confidence: 99%

“…Assays were performed as previously described 11 . pGL4-TOP reporter with TCF responsive elements to quantify ß-catenin transcriptional activity was a generous gift from Pr.…”

Section: Luciferase Reporter Assaysmentioning

confidence: 99%

Section: Western Blotmentioning

confidence: 99%

Section: Quantitative Real-time Pcrmentioning

confidence: 99%

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