Knock-Down of the 37-kDa/67-kDa Laminin Receptor in Mouse Brain by Transgenic Expression of Specific Antisense LRP RNA

Venous thrombosis occurs in patients with quantitative and qualitative fibrinogen disorders. Injury-induced thrombosis in zebrafish larvae has been used to model human coagulopathies. We aimed to determine whether zebrafish models of afibrinogenemia and dysfibrinogenemia have different thrombotic phenotypes. Laser injuries were used to induce venous thrombosis and the time-to-occlusion (TTO) and the binding and aggregation of fluorescent Tg(itga2b:EGFP) thrombocytes measured. The fga−/− larvae failed to support occlusive venous thrombosis and showed reduced thrombocyte binding and aggregation at injury sites. The fga+/− larvae were largely unaffected. When genome editing zebrafish to produce fibrinogen Aα R28C, equivalent to the human Aα R35C dysfibrinogenemia mutation, we detected in-frame skipping of exon 2 in the fga mRNA, thereby encoding AαΔ19–56. This mutation is similar to Fibrinogen Montpellier II which causes hypodysfibrinogenemia. Aα+/Δ19–56 fish had prolonged TTO and reduced thrombocyte activity, a dominant effect of the mutation. Finally, we used transgenic expression of fga R28C cDNA in fga knock-down or fga−/− mutants to model thrombosis in dysfibrinogenemia. Aα R28C expression had similar effects on TTO and thrombocyte activity as Aα+/Δ19–56. We conclude that thrombosis assays in larval zebrafish can distinguish between quantitative and qualitative fibrinogen disorder models and may assist in anticipating a thrombotic phenotype of novel fibrinogen mutations.

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Knock-Down of LRP/LR Influences Signalling Pathways in Late Stage Colorectal Carcinoma Cells

10.21203/rs.3.rs-74216/v1 ◽

2020 ◽

Author(s):

Leila Vania ◽

Gavin Morris ◽

Eloise Ferreira ◽

Stefan Weiss

Keyword(s):

Colorectal Cancer ◽

Cancer Cells ◽

Late Stage ◽

Telomerase Activity ◽

Laminin Receptor ◽

Signalling Pathways ◽

Continuous Growth ◽

Growth And Survival ◽

Knock Down ◽

Colorectal Cancer Cells

Abstract BackgroundThe 37kDa/67kDa laminin receptor (LRP/LR) is involved in several tumourigenic-promoting processes including cellular viability maintenance and apoptotic evasion. Thus, the aim of this study was to assess the molecular mechanism of LRP/LR on apoptotic pathways in late stage (DLD-1) colorectal cancer cells upon siRNA-mediated down-regulation of LRP/LR. MethodssiRNAs were used to down-regulate the expression of LRP/LR in DLD-1 cells which was assessed using western blotting and qPCR. To evaluate the mechanistic role of LRP/LR, proteomic analysis of pathways involved in proliferation and apoptosis were investigated. The data from the study was analysed using a one-way ANOVA, followed by a two-tailed student’s t-test with a confidence interval of 95%. ResultsHere we show that knock-down of LRP/LR led to significant changes in the proteome of DLD-1 cells, exposing new roles of the protein. Moreover, analysis showed that LRP/LR may alter components of the MAPK, p53-apoptotic and autophagic signalling pathways to aid colorectal cancer cells in continuous growth and survival. Knock-down of LRP/LR also resulted in significant decreases in telomerase activity and telomerase-related proteins in the DLD-1 cells. ConclusionsThese findings show that LRP/LR is critically implicated in apoptosis and cell viability maintenance and suggest that siRNA-mediated knock-down of LRP/LR may be a possible therapeutic strategy for the treatment of colorectal cancer.

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Different Isoforms of the Non-Integrin Laminin Receptor Are Present in Mouse Brain and Bind PrP

Biological Chemistry ◽

10.1515/bc.2003.027 ◽

2003 ◽

Vol 384 (2) ◽

Cited By ~ 28

Author(s):

S. Simoneau ◽

S. Haïk ◽

C. Leucht ◽

D. Dormont ◽

J.-P. Deslys ◽

...

Keyword(s):

Mouse Brain ◽

Laminin Receptor

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Knock-down of LRP/LR influences signalling pathways in late-stage colorectal carcinoma cells

BMC Cancer ◽

10.1186/s12885-021-08081-3 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Leila Vania ◽

Gavin Morris ◽

Eloise Ferreira ◽

Stefan F. T. Weiss

Keyword(s):

Colorectal Cancer ◽

Cancer Cells ◽

Late Stage ◽

Telomerase Activity ◽

Laminin Receptor ◽

Signalling Pathways ◽

Continuous Growth ◽

Growth And Survival ◽

Knock Down ◽

Colorectal Cancer Cells

Abstract Background The 37 kDa/67 kDa laminin receptor (LRP/LR) is involved in several tumourigenic-promoting processes including cellular viability maintenance and apoptotic evasion. Thus, the aim of this study was to assess the molecular mechanism of LRP/LR on apoptotic pathways in late stage (DLD-1) colorectal cancer cells upon siRNA-mediated down-regulation of LRP/LR. Methods siRNAs were used to down-regulate the expression of LRP/LR in DLD-1 cells which was assessed using western blotting and qPCR. To evaluate the mechanistic role of LRP/LR, proteomic analysis of pathways involved in proliferation and apoptosis were investigated. The data from the study was analysed using a one-way ANOVA, followed by a two-tailed student’s t-test with a confidence interval of 95%. Results Here we show that knock-down of LRP/LR led to significant changes in the proteome of DLD-1 cells, exposing new roles of the protein. Moreover, analysis showed that LRP/LR may alter components of the MAPK, p53-apoptotic and autophagic signalling pathways to aid colorectal cancer cells in continuous growth and survival. Knock-down of LRP/LR also resulted in significant decreases in telomerase activity and telomerase-related proteins in the DLD-1 cells. Conclusions These findings show that LRP/LR is critically implicated in apoptosis and cell viability maintenance and suggest that siRNA-mediated knock-down of LRP/LR may be a possible therapeutic strategy for the treatment of colorectal cancer.

Download Full-text