Myristoylated alanine‐rich C kinase substrate phosphorylation promotes cholangiocarcinoma cell migration and metastasis via the protein kinase C‐dependent pathway

Abstract

Myristoylated alanine-rich C kinase substrate (MARCKS), a substrate of protein kinase C (PKC) has been suggested to be implicated in cell adhesion, secretion, and motility through the regulation of the actin cytoskeletal structure. The quantitative real-time-polymerase chain reaction analysis revealed that MARCKS is significantly overexpressed in Opisthorchis viverriniassociated cholangiocarcinoma (CCA) (P = 0.001) in a hamster model, which correlated with the results of mRNA in situ hybridization. An immunohistochemical analysis of 60 CCA patients revealed a significant increase of MARCKS expression. Moreover, the log-rank analysis indicated that CCA patients with a high MARCKS expression have significantly shorter survival times than those with a low MARCKS expression (P = 0.02). This study investigated whether MARCKS overexpression is associated with CCA metastasis. Using a confocal microscopic analysis of CCA cell lines that had been stimulated with the PKC activator, 12-0-tetradecanoyl phorbol-13-acetate (TPA), MARCKS was found to be translocated from the plasma membrane to the perinuclear area. In addition, phosphorylated MARCKS (pMARCKS) became highly concentrated in the perinuclear area. Moreover, an adhesion assay demonstrated that the exogenous overexpression of MARCKS remarkably promoted cell attachment. Interestingly, after TPA stimulation, the CCA cell line-depleted MARCKS showed a decrease in migration and invasion activity. It can be concluded that in non-stimulation, MARCKS promotes cell attachment to the extracellular matrix. After TPA stimulation, PKC phosphorylates MARCKS leading to cell migration or invasion. Taken together, the results of this study reveal a prominent role for MARCKS as one of the key players in the migration of CCA cells and suggest that cycling between MARCKS and pMARCKS can regulate the metastasis of biliary cancer cells. (Cancer Sci 2010; 101: 658-665) C holangiocarcinoma (CCA), the malignant tumor arising from the bile duct epithelium, is the most common cancer in Thailand, especially in the northeast. There is strong epidemiological evidence (1)(2)(3) and experimental studies (4)(5)(6) that support an important etiological role for liver fluke (Opisthorchis viverrini [Ov]) infection in the development of CCA in humans. There is accumulating evidence that Ov infection enhances the risk of CCA via chronic inflammation. (7,8) Recently, Pinlaor et al. (9) suggested that oxidative and nitrosative damage to DNA in the liver of Ov-infected hamsters can play a key role in the modulation of gene expression, which enhances the effect of the DNA adduct and can drive a normal cell undergo tumor development. Additionally, Thanan et al. (10) reported that the highest 8-oxodG level, which is the biomarker of DNA damage, was observed for CCA patients, and a higher level was found in the urine and leukocytes of Ov-infected patients than that in healthy patients.The gene expression profiles of Ov-associated CCA tumors, as investigated by Loilome et al.(11) , indicated that myristoyla...

A Novel Predictor of Cancer Malignancy: Up-regulation of Myristoylated Alanine-Rich C Kinase Substrate Phosphorylation in Lung Cancer

“…This is because MARCKS expression is ubiquitous in various normal and tumor tissues. Despite this, there is a consensus that phospho-MARCKS, a post-translational modification, is associated with cell motility, and has a role in the regulation of cancer cell invasiveness and metastasis (2,4,14,15). Of note, our laboratory discovered that inhibition of MARCKS phosphorylation was able to reduce lung cancer metastasis in murine models (2).…”

Myristoylated Alanine‐Rich Protein Kinase Substrate (MARCKS) Regulates Small GTPase Rac1 and Cdc42 Activity and Is a Critical Mediator of Vascular Smooth Muscle Cell Migration in Intimal Hyperplasia Formation

“…MARCKS is a well‐established regulator of migration in multiple cell types. Dysregulation of MARCKS is closely associated with metastasis in a wide range of cancers and is an indicator of poor prognosis 19, 20. MARCKS increases cell motility through the protein kinase B pathway in fibroblasts21 and glial cells 22.…”

Over-expression of phosphorylated MARCKS in the nicotine-derived nitrosamine ketone induced lung cancer mice

“…The data confirms the hypothesis that phosphorylated MARCKS was overexpressed during lung cancer. To support the data, it was reported that the phosphorylated MARCKS specifically regulate the cancer migration and metastasis (Chen et al, 2014;Chen and Rotenberg, 2010;Ghoul et al, 2006;Reddy et al, 2010;Techasen et al, 2010).…”

“…At the same time it was also identified that in lungs, MARCKS especially phosphorylated MARCKS, plays a crucial role in controlling mucin secretion and inflammation (Eckert et al, 2010;Green et al, 2011;Li et al, 2001;Park et al, 2007;Singer et al, 2004). In addition, it was reported that phosphorylated MARCKS specifically regulate the cancer migration and metastasis (Chen et al, 2014;Chen and Rotenberg, 2010;Ghoul et al, 2006;Reddy et al, 2010;Techasen et al, 2010). In contrast, very little is known about the importance of phospho-MARCKS and its relevance in lung cancer.…”