The TLK1-MK5 axis regulates motility, invasion, and metastasis of prostate cancer cells

Abstract

ABSTRACTBackgroundMetastatic dissemination of prostate cancer (PCa) accounts for majority of PCa related deaths. However, the exact mechanism of PCa cell spread is still unknown. We uncovered a novel interaction between two unrelated promotility factors, tousled-like kinase 1 (TLK1) and MAPK-activated protein kinase 5 (MK5), which initiates a signaling cascade promoting metastasis. In PCa, TLK1-MK5 signaling might be crucial as androgen deprivation therapy (ADT) leads to increased expression of both TLK1 and MK5 in metastatic patients, but in this work, we directly investigated the motility, invasive, and metastatic capacity of PCa cells following impairment of the TLK1>MK5 axis.ResultsWe conducted scratch wound repair and 3D invasion assays with LNCaP and PC3 cells to determine if TLK1 and MK5 can regulate motility and invasion. Both genetic depletion and pharmacologic inhibition of TLK1 and MK5 resulted in reduced migration and invasion through a Matrigel plug. We further elucidated the potential mechanisms underlying these effects and found that that this is likely due to reorganization of the actin fibers at lamellipodia and the focal adhesions network, in conjunction with increased expression of some MMPs that can affect penetration through the ECM. PC3, a highly metastatic cell line when assayed in xenografts, was further tested in tail-vein injection/lung metastasis model, and we showed that following inoculation, treatment with GLPG (MK5 specific inhibitor) or J54 (TLK1 inhibitor) the resulting lung tumor nodules were greatly diminished in number, and for J54 also in size.ConclusionOur data support that TLK1-MK5 axis is functionally involved in driving PCa cell metastasis and clinical aggressiveness, hence, disruption of this axis may inhibit the metastatic capacity of PCa.SIMPLE SUMMARYRecent work by us and others has illustrated the critical importance of MK5/PRAK in the invasive and motility properties of several cancer cell lines and some mouse models. In our earlier work we also uncovered that TLK1 modulates the activity of MK5 by phosphorylating S354 and two additional sites (S160 and S386).. We have now expanded on the possible mechanisms of the TLK1>MK5 pro-motility and invasive activity, and report that this may be due to reorganization of the actin fibers at lamellipodia and the focal adhesions network, in conjunction with increased expression of some MMPs. Pharmacological or genetic manipulation of prostate cancer (PCa) cell lines, LNCaP and PC3, results in drastic loss of in vitro motility and invasive capacity of these cells concomitant with alterations of their general morphology and reorganization of the focal adhesions distribution. In addition, PC3 used in tail-vein experimental metastases studies show that the use of GLPG (MK5 inhibitor) or J54 (TLK1 inhibitor) results in a drastic reduction of metastatic lung nodules, macroscopically and histologically.