Identification of a Novel Human E‐Cadherin Splice Variant and Assessment of Its Effects Upon EMT‐Related Events

Matos, Matheus J. S.; Lapyckyj, Lara; Rosso, Marina; Besso, María José; Mencucci, María Victoria; Briggiler, Clara Isabel Marín; Giustina, Silvina; Furlong, Laura I.; Vazquez‐Levin, Mónica H.

doi:10.1002/jcp.25622

Cited by 35 publications

(40 citation statements)

References 81 publications

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“…EMT is characterized by switch E-cadherin to N-cadherin and Snail, thus greatly affecting the tumor metastasis [32,33,34]. As expected, we found that the knockdown of KIFC1 increased the expression of an E-cadherin, inhibited expression levels of EMT-related transcription factors N-cadherin, Snail and ZEB1, suggesting that KIFC1 is involved in the metastasis of PTC.…”

Section: Discussionsupporting

confidence: 84%

KIFC1 accelerates the proliferation, migration and invasion of papillary thyroid cancer cells via MAPK signaling

Nie¹,

Han²,

Wen³

et al. 2020

Preprint

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Kinesin family member C1 (KIFC1) acts as a kind of minus end-directed motorized protein and is considered as an oncogene of some cancer types. However, no studies have fully elucidated its biological activity and molecular mechanisms in papillary thyroid cancer (PTC). The study focused on reporting the overexpression of KIFC1 in cell lines and tissues of PTC. Moreover, clinicopathological features analysis showed that KIFC overexpression is significantly correlated with extrathyroidal invasion and lymph node metastasis. Knockdown of KIFC1 significantly reduced cell growth, migration and invasion in PTC cells, and concomitant increased levels of differentiation markers, such as Tg and Nis. Knockdown of KIFC1 markedly increased the expression level of epithelial cell marker (E-cadherin), and decreased the expression levels of epithelial-mesenchymal transition (EMT) related transcriptional factor N-cadherin, Snail and ZEB1. Further study revealed that knockdown of KIFC1 downregulated stemness markers ALDH2 and SOX2, and inhibited the MAPK signaling cascades and downstream signaling, including p-ERK, ERK, p-JNK, JNK, MMP2, and MMP9, which can affect the expression of the EMT associated factors. Taken together, we reported that KIFC1 might promoted the proliferation, migration and invasion of PTC cells and offer a candidate molecular target for therapeutic intervention.

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Section: Discussionsupporting

confidence: 84%

KIFC1 accelerates the proliferation, migration and invasion of papillary thyroid cancer cells via MAPK signaling

Nie¹,

Han²,

Wen³

et al. 2020

Preprint

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“…We hypothesise that this extensive unproductive splicing may participate to the regulation of the expression levels of PKHD1, through a mechanism of gene regulation known as alternative splicing-coupled nonsense-mediated decay (AS-NMD) (42)(43)(44). Disturbance to this process, induced by splicing variants such as c.6900C>T p.(Asn2300=), can contribute to the pathogenicity of ARPKD, as previously shown for cancer and autoimmune diseases (45)(46)(47). The synonymous variant p. (Asn2300=), by promoting the production of the premature stop codon-containing transcript B, may decrease the levels of PKHD1 expression below a certain threshold required for its normal function.…”

Section: Discussionmentioning

confidence: 76%

Use of patient derived urine renal epithelial cells to confirm pathogenicity of PKHD1 alleles

Molinari

Srivast

Dewhurst

et al. 2020

Preprint

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Background PKHD1 is the main genetic cause of autosomal recessive polycystic kidney disease (ARPKD), a hereditary hepato-renal fibrocystic disorder which is the most important cause of end-stage renal disease during early childhood. ARPKD can also present in adulthood with milder phenotypes. In this study, we describe a 24-year-old woman with atypical polycystic kidney, no family history of renal disease and no obvious extra-renal manifestations who was referred for genetic investigation. Methods We used a combination of next generation sequencing, Sanger sequencing and RNA and microscopy studies performed on urine-derived renal epithelial cells (URECs) to provide a genetic diagnosis of ARPKD. Results A next generation sequencing panel of cystic ciliopathy genes allowed the identification of two heterozygous sequence changes in PKHD1 (c.6900C>T; p.(Asn2300=) and c.7964A>C; p.(His2655Pro)). The pathogenicity of the synonymous PKHD1 variant is not clear and requires RNA studies, which cannot be carried out efficiently on RNA extracted from proband blood, due to the low expression levels of PKHD1 in lymphocytes. Using URECs as a source of kidney-specific RNA, we show that PKHD1 is alternatively spliced around exon 43, both in control and proband URECs. The variant p.(Asn2300=) shifts the expression ratio in favour of a shorter, out-of-frame transcript. To further study the phenotypic consequence of these variants, we investigated the ciliary phenotype of patient URECs, which were abnormally elongated and presented multiple blebs along the axoneme. Conclusions We confirm the power of URECs as a tool for functional studies on candidate variants in inherited renal disease, especially when the expression of the gene of interest is restricted to the kidney and we describe, for the first time, ciliary abnormalities in ARPKD patient cells.

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“…MB49-I cells transient transfection was performed using Lipofectamine R 2000 (Thermo) and 100 pmol/ml of a specific small interfering Ephrin-B1 RNA (siRNA) (#39437, SCB; pool of 3 target-specific 19-25 nt siRNAs designed to knock down gene expression) following a standard procedure (18). Control assays with scramble siRNA (#37007, SCB) were included.…”

Section: Transient Transfection Assaysmentioning

confidence: 99%

“…RNA Extraction, cDNA Synthesis, Standard, and Quantitative Real Time PCR Cell and tissue total RNA was isolated and processed for complementary DNA (cDNA) synthesis (14,15,18). PCR samples were prepared with SYBR Green R PCR Master Mix (Thermo) mixture and run in CFX96 Touch TM…”

Section: Transient Transfection Assaysmentioning

confidence: 99%

“…Protein extraction and analysis was done essentially as previously described (14,15,18). Basically, cell monolayers were incubated with PBS on ice, followed by protein extraction with RIPA (20 mM Tris-HCl, pH = 7.5, 150 mM NaCl, 1% NP-40, 1% sodium deoxycholate, and standard protease inhibitors cocktail) buffer.…”

Section: Protein Extraction and Western Immunoblotting Protocolsmentioning

confidence: 99%

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Ephrin-B1 Is a Novel Biomarker of Bladder Cancer Aggressiveness. Studies in Murine Models and in Human Samples

et al. 2020

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Bladder cancer (BC) is the ninth most common cancer worldwide, but molecular changes are still under study. During tumor progression, Epithelial cadherin (E-cadherin) expression is altered and β-catenin may be translocated to the nucleus, where it acts as co-transcription factor of tumor invasion associated genes. This investigation further characterizes E-cadherin and β-catenin associated changes in BC, by combining bioinformatics, an experimental murine cell model (MB49/MB49-I) and human BC samples. In in silico studies, a DisGeNET (gene-disease associations database) analysis identified CDH1 (E-cadherin gene) as one with highest score among 130 BC related-genes. COSMIC mutation analysis revealed CDH1 low mutations rates. Compared to MB49 control BC cells, MB49-I invasive cells showed decreased E-cadherin expression, E-to P-cadherin switch, higher β-catenin nuclear signal and lower cytoplasmic p-Ser33-β-catenin signal, higher Ephrin-B1 ligand and EphB2 receptor expression, higher Phospho-Stat3 and Urokinase-type Plasminogen Activator (UPA), and UPA receptor expression. MB49-I cells transfected with Ephrin-B1 siRNA showed lower migratory and invasive capacity than control cells (scramble siRNA). By immunohistochemistry, orthotopic MB49-I tumors had lower E-cadherin, increased nuclear β-catenin, lower pSer33-β-catenin cytoplasmic signal, and higher Ephrin-B1 expression than MB49 tumors. Similar changes were found in human BC tumors, and 83% of infiltrating tumors depicted a high Ephrin-B1 stain. An association between higher Ephrin-B1 expression and higher stage and tumor grade was found. No association was found between abnormal E-cadherin signal, Ephrin-B1 expression or clinical-pathological parameter. This study thoroughly analyzed E-cadherin and associated changes in BC, and reports Ephrin-B1 as a new marker of tumor aggressiveness.

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Identification of a Novel Human E‐Cadherin Splice Variant and Assessment of Its Effects Upon EMT‐Related Events

Cited by 35 publications

References 81 publications

KIFC1 accelerates the proliferation, migration and invasion of papillary thyroid cancer cells via MAPK signaling

KIFC1 accelerates the proliferation, migration and invasion of papillary thyroid cancer cells via MAPK signaling

Use of patient derived urine renal epithelial cells to confirm pathogenicity of PKHD1 alleles

Ephrin-B1 Is a Novel Biomarker of Bladder Cancer Aggressiveness. Studies in Murine Models and in Human Samples

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