Cell Cycle–Dependent Tumor Engraftment and Migration Are Enabled by Aurora-A

Abstract

Cell-cycle progression and the acquisition of a migratory phenotype are hallmarks of human carcinoma cells that are perceived as independent processes but may be interconnected by molecular pathways that control microtubule nucleation at centrosomes. Here, cell-cycle progression dramatically impacts the engraftment kinetics of 4T1-luciferase2 breast cancer cells in immunocompetent BALB/c or immunocompromised NOD-SCID gamma (NSG) mice. Multiparameter imaging of wound closure assays was used to track cell-cycle progression, cell migration, and associated phenotypes in epithelial cells or carcinoma cells expressing a fluorescence ubiquitin cell-cycle indicator. Cell migration occurred with an elevated velocity and directionality during the S-G-phase of the cell cycle, and cells in this phase possess front-polarized centrosomes with augmented microtubule nucleation capacity. Inhibition of Aurora kinase-A (AURKA/Aurora-A) dampens these phenotypes without altering cell-cycle progression. During G-phase, the level of phosphorylated Aurora-A at centrosomes is reduced in hyaluronan-mediated motility receptor (HMMR)-silenced cells as is the nuclear transport of TPX2, an Aurora-A-activating protein. TPX2 nuclear transport depends upon HMMR-T703, which releases TPX2 from a complex with importin-α (KPNA2) at the nuclear envelope. Finally, the abundance of phosphorylated HMMR-T703, a substrate for Aurora-A, predicts breast cancer-specific survival and relapse-free survival in patients with estrogen receptor (ER)-negative ( = 941), triple-negative (TNBC) phenotype ( = 538), or basal-like subtype ( = 293) breast cancers, but not in those patients with ER-positive breast cancer ( = 2,218). Together, these data demonstrate an Aurora-A/TPX2/HMMR molecular axis that intersects cell-cycle progression and cell migration. Tumor cell engraftment, migration, and cell-cycle progression share common regulation of the microtubule cytoskeleton through the Aurora-A/TPX2/HMMR axis, which has the potential to influence the survival of patients with ER-negative breast tumors. .

Hyaluronan Mediated Motility Receptor (HMMR) Encodes an Evolutionarily Conserved Homeostasis, Mitosis, and Meiosis Regulator Rather than a Hyaluronan Receptor

“…While full-length HMMR is able to locate to centrosomes and bind to microtubules [16], the naturally occurring splice variant HMMR (-exon 4) localizes to the mitotic spindle during mitosis but shows nuclear localization during interphase [16,19]. Localization to the nucleus is also seen for a phosphorylated form of HMMR at Thr703 [22]. Phosphorylation at Thr703 was identified by phospho-proteomics [23] and, through the use of antibodies specific for phospho-Thr703 HMMR, it was revealed that phospho-Thr703 not only directs the protein to the nucleus but also may assist in the regulation of the Ran-dependent nuclear transport of targeting protein for Xklp2 (TPX2) [22].…”

“…Localization to the nucleus is also seen for a phosphorylated form of HMMR at Thr703 [22]. Phosphorylation at Thr703 was identified by phospho-proteomics [23] and, through the use of antibodies specific for phospho-Thr703 HMMR, it was revealed that phospho-Thr703 not only directs the protein to the nucleus but also may assist in the regulation of the Ran-dependent nuclear transport of targeting protein for Xklp2 (TPX2) [22].…”

“…In addition to such hypothesis-based studies, a number of non-biased, genome-wide analyses also indicate similar mitotic roles of HMMR. The All RNA-Seq and Chip-Seq Sample and Signature Search (ARCHS4) tool, which mines sequencing data from 103,083 mouse and 84,863 human samples [56], predicts HMMR functions in processes related to mitosis and chromosome segregation, kinetochore, and nuclear pore complex assembly, which have all been independently and experimentally validated [16,22,35]. Using this tool [56], HMMR is found to be co-expressed with TOP2A, KIF11, TPX2, ECT2, BUB1, KIF20A, NUSAP1, KIF20B, SMC2, and CCNA2; each of these gene products are involved with cell cycle regulation, spindle organization, and chromosome segregation.…”

Curcuma zanthorrhiza Extracts Induce G2/M Cell Cycle Arrest and Apoptosis in 4T1 and MCF-7 Human Breast Cancer Cells

“…This inability is related to the effect of temulawak in inducing cell cycles arrest in the G2/M phase. Chu et al, 2018, reported that cellcycle progression and the migratory acquisition are found as independent processes but may be interconnected. The interconnection is through molecular pathways that control microtubule nucleation at centrosomes.…”

Examining go-or-grow using fluorescent cell-cycle indicators and cell cycle-inhibiting drugs

“…1(a)). FUCCI allows us to study cell motility in G1 separately from cell motility in S/G2/M [6,20,26,27]. Specifically, we investigate cycling cells for differences in motility when the cells are in G1 compared with S/G2/M.…”