Golgi Stabilization, Not Its Front-Rear Bias, Is Associated with EMT-Enhanced Fibrillar Migration

Abstract

Epithelial-to-mesenchymal transition (EMT) and maturation of collagen fibrils in the tumor microenvironment play a significant role in cancer cell invasion and metastasis. Confinement along fiber-like tracks enhances cell migration. To what extent and in what manner EMT further promotes migration in a microenvironment already conducive to migration is poorly understood. Here, we show that TGFb-mediated EMT significantly enhances migration on fiber-like micropatterned tracks of collagen, doubling migration speed and tripling persistence relative to untreated mammary epithelial cells. Thus, cell-intrinsic EMT and extrinsic fibrillar tracks have nonredundant effects on motility. To better understand EMT-enhanced fibrillar migration, we investigated the regulation of Golgi positioning, which is involved in front-rear polarization and persistent cell migration. Confinement along fiber-like tracks has been reported to favor posterior Golgi positioning, whereas anterior positioning is observed during 2-day wound healing. Although EMT also regulates cell polarity, little is known about its effect on Golgi positioning. Here, we show that EMT induces a 2:1 rearward bias in Golgi positioning; however, positional bias explains less than 2% of single-cell variability in migration speed and persistence. Meanwhile, EMT significantly stabilizes Golgi positioning. Cells that enhance migration in response to TGFb maintain Golgi position for 2-to 4-fold longer than nonresponsive counterparts irrespective of whether the Golgi is ahead or behind the nucleus. In fact, 28% of TGFb-responsive cells exhibit a fully committed Golgi phenotype with the organelle either in the anterior or posterior position for over 90% of the time. Furthermore, single-cell differences in Golgi stability capture up to 18% of variations in migration speed. These results suggest a hypothesis that the Golgi may be part of a core physical scaffold that affects how cell-generated forces are distributed during migration. A stable scaffold would be expected to more consistently and therefore more productively distribute forces over time, leading to efficient migration.

Stimulation of cell invasion by the Golgi Ion Channel GAAP/TMBIM4 via an H2O2-Dependent Mechanism

“…Whilst various reports have recognized the involvement of the GA in regulating cell invasion, this is the first report of a GA resident ion channel controlling cell invasion and mitochondrial metabolic status. The positioning of the GA at the front of migrating cells mediates the delivery of new membrane and adhesion proteins to the leading edge and is closely linked with cell polarity and directional migration [[65], [66], [67]]. Several GA proteins have been linked with cell migration and are associated with cancer progression.…”

Study on Targeting Relationship Between miR-320b and FGD5-AS1 and Its Effect on Biological Function of Osteosarcoma Cells

“…Changes in the biological traits of EMT will result in cells acquiring the ability to migrate and invade, while maintaining stem cell characteristics, tumor progression, and metastasis are all related to EMT. 27,28 By detecting EMT-related proteins, we observed that FGD5-AS1 inhibition led to decreased EMT behavior, increased E-cadherin, and decreased N-cadherin (P<0.05), suggesting that FGD5-AS1 could be used as a therapeutic target for osteosarcoma to regulate the malignant behavior of tumor cells. Increasing evidence has demonstrated that lncRNAs, highly expressed in the cytoplasm, can act as molecular sponges, competitively bind or adsorb with other regulatory proteins and miRNAs, thus participating in the biological processes of tumors.…”

Multivariate relationships among nucleus and Golgi properties during fibrillar migration are robust to and unchanged by epithelial-to-mesenchymal transition

“…Automated segmentation of time-lapse images of fluorescently labeled migrating cells was completed in MATLAB using a custom program described in prior work [15]. Briefly, the regionprops function was used to calculate the area, perimeter, and centroid of the nucleus and Golgi.…”

“…Nucleus orientation (12) refers to nucleus orientation with respect to direction of migration. Asterisk denotes properties that were calculated and reported in [15].…”

“…However, when cells are confined to migrate along narrow fiber-like micropatterns to mimic a fibrillar collagen microenvironment, the Golgi is positioned posterior to the nucleus [ 13 , 14 ]. We have recently shown that more significant than the position of the Golgi is the stability with which its position is maintained [ 15 ]. The stability of Golgi positioning, not whether the Golgi is ahead or behind the nucleus, is associated with EMT-mediated enhancement of fibrillar migration.…”