Effect of LIM kinase 1 overexpression on behaviour of endometriosis-derived stromal cells

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LIM kinase 1 is overexpressed in endometriosis-derived stromal cells, promoting their malignant-like behaviors including migration, invasion, proliferation, and angiogenesis.

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The study examined whether LIM kinase 1 (LIMK1) is differently expressed in endometriosis-derived stromal cells and tested how altering LIMK1 levels affects their in vitro behavior. Endometriosis patients (n=30) provided eutopic endometrial stromal cells (ESCs) and non-endometriosis patients (n=30) provided normal endometrial stromal cells (NSCs); LIMK1 mRNA and protein were quantified by real-time PCR and Western blot, then LIMK1 was silenced in ESCs using lentiviral siRNA and overexpressed in NSCs using lentiviral LIMK1 cDNA. ESCs had higher basal LIMK1 expression and showed greater migration, invasion, proliferation, and higher adhesion/invasion/angiogenesis marker expression than NSCs, while LIMK1 knockdown reduced ESC malignant-like behavior and LIMK1 upregulation increased NSC malignant-like behavior. A key caveat is that findings are limited to in vitro assays in stromal cells rather than in vivo validation. This paper is centrally about endometriosis—investigating LIMK1 overexpression and its effects on malignant-like migration, invasion, proliferation, and angiogenesis in endometriosis-derived stromal cells.

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Abstract

LIM kinase 1 (LIMK1) has been implicated in tumour invasion and migration in several tumour types. However, its role in the progression of endometriosis has not yet been studied. Our aim is to analyse LIMK1 expression in endometriosis-derived stromal cells and to explore the effects of LIMK1 overexpression on their biological behaviour. The mRNA and protein expression levels of LIMK1 of eutopic endometrial stromal cells (ESCs) separated from 30 endometriosis patients and normal endometrial stromal cells (NSCs) separated from 30 patients without endometriosis were analysed by real-time polymerase chain reaction and Western blot. Lentiviral particles containing human LIMK1 short interfering RNA were used to silence the LIMK1 gene in ESCs and LIMK1-expressing lentiviral particles containing the total LIMK1 cDNA was transfected into NSCs to upregulate LIMK1 expression. Cell migration, invasion, proliferation and expression of markers of adhesion, invasion and angiogenesis of ESCs and NSCs were evaluated under basal conditions and after transfection in vitro. The mRNA and protein expression levels of LIMK1 in ESCs were higher than those in NSCs. Under basal conditions, ESCs exhibited greater cell migration, invasion and proliferation and higher levels of markers of adhesion, invasion and angiogenesis than NSCs. The behaviour of ESCs was decreased after LIMK1 gene silencing and that of NSCs was elevated after LIMK1 gene upregulation. Thus, LIMK1 is overexpressed in ESCs thereby facilitating malignant-like behaviour, including enhanced migration, invasion, proliferation and angiogenesis, all of which contribute to the occurrence and development of endometriosis.
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Abstract

LIM kinase 1 (LIMK1) has been implicated in tumour invasion and migration in several tumour types. However, its role in the progression of endometriosis has not yet been studied. Our aim is to analyse LIMK1 expression in endometriosis-derived stromal cells and to explore the effects of LIMK1 overexpression on their biological behaviour. The mRNA and protein expression levels of LIMK1 of eutopic endometrial stromal cells (ESCs) separated from 30 endometriosis patients and normal endometrial stromal cells (NSCs) separated from 30 patients without endometriosis were analysed by real-time polymerase chain reaction and Western blot. Lentiviral particles containing human LIMK1 short interfering RNA were used to silence the LIMK1 gene in ESCs and LIMK1-expressing lentiviral particles containing the total LIMK1 cDNA was transfected into NSCs to upregulate LIMK1 expression. Cell migration, invasion, proliferation and expression of markers of adhesion, invasion and angiogenesis of ESCs and NSCs were evaluated under basal conditions and after transfection in vitro. The mRNA and protein expression levels of LIMK1 in ESCs were higher than those in NSCs. Under basal conditions, ESCs exhibited greater cell migration, invasion and proliferation and higher levels of markers of adhesion, invasion and angiogenesis than NSCs. The behaviour of ESCs was decreased after LIMK1 gene silencing and that of NSCs was elevated after LIMK1 gene upregulation. Thus, LIMK1 is overexpressed in ESCs thereby facilitating malignant-like behaviour, including enhanced migration, invasion, proliferation and angiogenesis, all of which contribute to the occurrence and development of endometriosis. Similar content being viewed by others

References

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Effect of LIM kinase 1 overexpression on behaviour of endometriosis-derived stromal cells. Cell Tissue Res 359, 885–893 (2015). https://doi.org/10.1007/s00441-014-2068-5 Received: Accepted: Published: Issue date: DOI: https://doi.org/10.1007/s00441-014-2068-5

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endometriosis

MeSH descriptors

Endometriosis Endometriosis Lim Kinases Adult Cell Movement Cell Proliferation Endometriosis Female Gene Knockdown Techniques Gene Silencing Humans Intercellular Adhesion Molecule-1 Intercellular Adhesion Molecule-1 Lim Kinases Matrix Metalloproteinase 9 Matrix Metalloproteinase 9 Stromal Cells Stromal Cells Stromal Cells Transfection

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