Activation of Vimentin Is Critical to Promote a Metastatic Potential of Cholangiocarcinoma Cells

Abstract

Cholangiocarcinoma (CCA) is a highly metastatic tumor, and the majority of patients with CCA have a short survival time because there are no available effective treatments. Hence, a better understanding regarding CCA metastasis may provide an opportunity to improve the strategies for treatment. A comparison study between the highly metastatic cells and their parental cells is an approach to uncover the molecular mechanisms underlying the metastatic process. In the present study, a lung metastatic CCA cell line, KKU-214L5, was established by the in vivo selection of the tail vein-injected mouse model. KKU-214L5 cells possessed mesenchymal spindle-like morphology with higher migration and invasion abilities in vitro than the parental cells (KKU-214). KKU-214L5 also exhibited extremely aggressive lung colonization in the tail vein-injected metastatic model. Epithelial-mesenchymal transition (EMT) was clearly observed in KKU-214L5 cells. Significant downregulation of epithelial markers (ZO-1 and claudin-1), with unique upregulation of E-cadherin and mesenchymal markers (vimentin, β-catenin, and slug), was observed in KKU-214L5. Increasing MMP-2 and MMP-9 activities and CD147 expression reflected the high invasion activity in KKU-214L5 cells. Suppression of vimentin using siRNA significantly decreased the migration and invasion capabilities of KKU-214L5 to almost the basal levels of the parental cells without any change on the expression levels of other EMT markers and the activities of MMPs. These results suggest that vimentin activation is essential to potentiate the metastatic characters of CCA cells, and suppression of vimentin expression could be a potential strategy to improve the treatment of CCA, a highly metastatic cancer.

A comparative study of the neural stem cell niche in the adult hypothalamus of human, mouse, rat and gray mouse lemur (Microcebus murinus)

“…The M0725 antibody from Dako detects a band at 54 kDa on Western blots of human mesenchymal stromal cell lysates (Talele, Fradette, Davies, Kapus, & Hinz, ). Treatment of a human cholangiocarcinoma cell line with siRNA against vimentin suppressed immunoblot and immunofluorescent signals with this antibody (Saentaweesuk et al, ). The two anti‐vimentin antibodies used yielded a similar expression profile.…”

Allicin Inhibits Proliferation and Invasion in Vitro and in Vivo via SHP-1-Mediated STAT3 Signaling in Cholangiocarcinoma

“…During the process of EMT, carcinoma cells lose their epithelial characteristics, including polarity and cell-cell adhesions, and acquire a mesenchymal phenotype to obtain an invasive capacity [50]. Several studies have indicated that the expression of MMP-2 and MMP-9 is associated with EMT, and vimentin is often used as a marker of mesenchymally derived cells or cells undergoing EMT during both normal development and metastatic progression [51,52]. However, the STAT3 signaling pathway has been shown to be involved in tumor EMT [53].…”

Increase of MAL-II Binding Alpha2,3-Sialylated Glycan Is Associated with 5-FU Resistance and Short Survival of Cholangiocarcinoma Patients

“…CCA cell lines, KKU-213 and KKU-214, were established from a primary tumor of CCA patients and deposited in JCRB. KKU-213L5 and KKU-214L5 were the lung metastatic CCA cell lines derived from KKU-213 and KKU-214 as previously described [27,28]. All cell lines were cultured in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% heat-inactivated fetal bovine serum (FBS) and antibiotic-antimycotic in a 5% CO 2 incubator at 37 • C.…”