M3 muscarinic acetylcholine receptors regulate epithelial–mesenchymal transition, perineural invasion, and migration/metastasis in cholangiocarcinoma through the AKT pathway

Abstract

BackgroundCholangiocarcinoma is a highly malignant tumor type that is not sensitive to radiotherapy or chemotherapy due to aggressive perineural invasion and metastasis. Unfortunately, the mechanisms underlying these processes and the signaling factors involved are largely unknown. In this study, we analyzed the role of M3 muscarinic acetylcholine receptors (M3-mAChR) in cell migration, perineural invasion, and metastasis during cholangiocarcinoma.MethodsWe assessed 60 human cholangiocarcinoma tissue samples and 30 normal biliary tissues. Immunohistochemical staining was used to detect M3-mAChR expression and the relationship between expression and clinical prognosis was evaluated. The biological functions of M3-mAChR in cholangiocarcinoma cell migration, perineural invasion, and epithelial–mesenchymal transition (EMT) were investigated using the human cholangiocarcinoma cell lines FRH0201 and RBE in conjunction with various techniques, including agonist/antagonist treatment, RNA interference, M3-mAChR overexpression, dorsal root ganglion co-culturing, immunohistochemistry, western blotting, etc.ResultsM3-mAChR were highly expressed in cholangiocarcinoma tissue and expression was closely related to differentiation and lymphatic metastasis, affecting patient survival. Treatment with the M3-mAChR agonist pilocarpine and M3-mAChR overexpression significantly promoted migration and perineural invasion, while the M3-mAChR antagonist atropine blocked these effects. Similarly, M3-mAChR knock-down also weakened cell migration and perineural invasion. The expression of phosphatase and tensin homolog, AKT, E-cadherin, vimentin, and Snail, which are components of the phosphatidylinositol 3-kinase/AKT signaling pathway and EMT, were altered by pilocarpine, and these effects were again blocked by atropine. Notably, AKT knock-down decreased M3-mAChR expression and reversed the downstream effects of this receptor.ConclusionsM3-mAChR are involved in tumor cell migration, perineural invasion, and EMT during cholangiocarcinoma, and these effects are modulated via the AKT signaling pathway.

<p>Perineural Invasion and Postoperative Complications are Independent Predictors of Early Recurrence and Survival Following Curative Resection of Gastric Cancer</p>

“…On the one hand, nerve tissue promotes tumor proliferation and invasion through specific signals; on the other hand, tumor cells can lead to the formation and extension of nerve cell axons and make nerve fibers thicker, which is conducive to the contact, adhesion and migration of tumor cells and nerves. [18][19][20] A meta-analysis including 30,590 gastric cancer patients who had undergone curative gastrectomy from 24 studies showed that the median rate of positive perineural invasion was 40.9% (6.8-75.6%). 21 The positivity rate of PNI increased when tumors were undifferentiated and the depth of invasion, the size, and clinical stage of tumor increased.…”

Small molecule inhibitors of epithelial‐mesenchymal transition for the treatment of cancer and fibrosis

“…For example, tripartite motif‐containing 14 is an oncogene that regulated EMT and the metastasis of human gastric cancer by activating Akt signaling . M3 muscarinic acetylcholine receptors regulated EMT, perineural invasion, and metastasis in cholangiocarcinoma via the Akt pathway . Canopy homologue 2 promoted EMT via activating the Akt/GSK3 pathway in non–small‐cell lung cancer .…”

“…168 M3 muscarinic acetylcholine receptors regulated EMT, perineural invasion, and metastasis in cholangiocarcinoma via the Akt pathway. 169 Canopy homologue 2 promoted EMT via activating the Akt/GSK3 pathway in non-small-cell lung cancer. 170 The Akt signaling pathway mediated cigarette-induced EMT in lung cancer.…”

A Simple Competing Endogenous RNA Network Identifies Novel mRNA, miRNA, and lncRNA Markers in Human Cholangiocarcinoma

“…Cholangiocarcinoma (CCA), which emerges from the malignant proliferation of cholangiocytes, is an epithelial tumor of the biliary trees [24]. Concealed symptoms, a lack of early diagnostic markers, and low sensitivity to conventional radiotherapy and chemotherapy treatments are the crucial causes underlying poor prognoses for CCA patients [25]. Due to high morbidity and mortality in CCA, revealing the underlying molecular mechanisms, identifying molecular biomarkers for early diagnosis, prevention, and personalized therapy are critically important and highly demanded.…”