LIMD2 Regulates Key Steps of Metastasis Cascade in Papillary Thyroid Cancer Cells via MAPK Crosstalk

Abstract

Although papillary thyroid carcinoma (PTC) has a good prognosis, 20–90% of patients show metastasis to regional lymph nodes and 10–15% of patients show metastasis to distant sites. Metastatic disease represents the main clinical challenge that impacts survival rate. We previously showed that LIMD2 was a novel metastasis-associated gene. In this study, to interrogate the role of LIMD2 in cancer invasion and metastasis, we used CRISPR-mediated knockout (KO) of LIMD2 in PTC cells (BCPAP and TPC1). Western blot and high-content screening (HCS) analysis confirmed functional KO of LIMD2. LIMD2 KO reduced in vitro invasion and migration. Ultrastructural analyses showed that cell polarity and mitochondria function and morphology were restored in LIMD2 KO cells. To unveil the signals supervising these phenotypic changes, we employed phospho-protein array. Several members of the MAPK superfamily showed robust reduction in phosphorylation. A Venn diagram displayed the overlap of kinases with reduced phosphorylation in both cell lines and showed that they were able to initiate or sustain the epithelial-mesenchymal transition (EMT) and DNA damage checkpoint. Flow cytometry and HCS validation analyses further corroborated the phospho-protein array data. Collectively, our findings show that LIMD2 enhances phosphorylation of kinases associated with EMT and invasion. Through cooperation with different kinases, it contributes to the increased genomic instability that ultimately promotes PTC progression.

LIMD2 Is a Prognostic and Predictive Marker in Patients With Esophageal Cancer Based on a Cerna Network Analysis

“…The survival time of NSCLC patients with overexpression of LIMD2 was shorter than that of patients with lower expression, indicating that LIMD2 may serve as a therapeutic target in NSCLC [45]. Araldi et al found that LIMD2 may strengthen phosphorylation of kinases related to epithelial-mesenchymal transition (EMT) and invasion, and modulate the crucial steps in the metastatic cascade of thyroid cancer cells through the mitogen-activated protein kinase (MAPK) crosstalk [47].…”

Exosomes in the Tumor Microenvironment: From Biology to Clinical Applications

“…Metastasis is a multistep process, which involves local invasion, intravasation, transport, extravasation, and colonization [88]. These steps require a series of genetic, biochemical, and morphological deregulations that are present in an evolutionarily conserved developmental program known as the epithelial-mesenchymal transition (EMT) [64,[89][90][91].…”

“…During embryogenesis, the EMT (EMT type I) gives rise to mesoderm (responsible for the formation of muscle, bone, and connective tissues) during gastrulation and neural crest delamination (which results in glial cell, adrenal gland, and epithelial pigmented cell formation) [90,96]. In adult life, the EMT plays a key role in tissue re-epithelization during wound healing (EMT type II) [95,97,98] but, when inappropriately active, such as occurs in carcinogenesis (EMT type III), the EMT causes important disturbances in epithelial tissue homeostasis and integrity, leading to cancer cell spread and metastasis [89,99].…”

“…HIF-1α nuclear translocation promotes the upregulation and stabilization of Snail and Twist, resulting in cadherin switching, which is characterized by the downregulation of E-cadherin (leading to a loss of intercellular adhesion and consequent activation of the Wnt/β-catenin pathway) and N-cadherin upregulation in cancer cells [103][104][105]. Combined with the F-actin reorganization of invadopodia sites, these actions create sites of transient adhesion that confer cell motility, facilitating the dissemination of cancer cells [89,106].…”

“…Cumulative evidence has shown that genomic instability contributes to CSC formation and accelerates the development of many genetically variable cancer stem cells, increasing the intratumor heterogeneity [89,[184][185][186][187].…”