RT Journal Article
T1 Depletion of ALMS1 affects TGF-β signalling pathway and downstream processes such as cell migration and adhesion capacity
A1 Bea Mascato, Brais
A1 Neira Goyanes, Elena
A1 Iglesias Rodríguez, Antía
A1 Valverde Pérez, Diana
K1 2410.07 Genética Humana
K1 3201.02 Genética Clínica
K1 2407.02 Citogenética
AB Background: ALMS1 is a ubiquitous gene associated with Alström syndrome(ALMS). The main symptoms of ALMS affect multiple organs and tissues,generating at last, multi-organic fibrosis in the lungs, kidneys and liver. TGF-β is one of the main pathways implicated in fibrosis, controlling the cell cycle,apoptosis, cell migration, cell adhesion and epithelial-mesenchymal transition(EMT). Nevertheless, the role of ALMS1 gene in fibrosis generation and otherimplicated processes such as cell migration or cell adhesion via the TGF- βpathway has not been elucidated yet.Methods: Initially, we evaluated how depletion of ALMS1 affects differentprocesses like apoptosis, cell cycle and mitochondrial activity in HeLa cells.Then, we performed proteomic profiling with TGF-β stimuli in HeLa ALMS1 −/−cells and validated the results by examining different EMT biomarkers usingqPCR. The expression of these EMT biomarkers were also studied in hTERT-BJ-5ta ALMS1 −/−. Finally, we evaluated the SMAD3 and SMAD2 phosphorylationand cell migration capacity in both models.Results: Depletion of ALMS1 generated apoptosis resistance to thapsigargin(THAP) and C2-Ceramide (C2-C), and G2/M cell cycle arrest in HeLa cells. Formitochondrial activity, results did not show significant differences betweenALMS1 +/+ and ALMS1 −/−. Proteomic results showed inhibition of downstreampathways regulated by TGF-β. The protein-coding genes (PCG) were associatedwith processes like focal adhesion or cell-substrate adherens junction in HeLa.SNAI1 showed an opposite pattern to what would be expected when activatingthe EMT in HeLa and BJ-5ta. Finally, in BJ-5ta model a reduced activation ofSMAD3 but not SMAD2 were also observed. In HeLa model no alterations in thecanonical TGF-β pathway were observed but both cell lines showed a reductionin migration capacity.Conclusion: ALMS1 has a role in controlling the cell cycle and the apoptosisprocesses. Moreover, the depletion of ALMS1 affects the signal transductionthrough the TGF-β and other processes like the cell migration and adhesioncapacity.
PB Frontiers in Molecular Biosciences
SN 2296889X
YR 2022
FD 2022-10-13
LK http://hdl.handle.net/11093/4411
UL http://hdl.handle.net/11093/4411
LA eng
NO Frontiers in Molecular Biosciences, 9, 9:992313 (2022)
NO Ministerio de Educación, Cultura y Deporte | Ref. FPU17/01567
DS Investigo
RD 13-oct-2024