MicroRNA-128 Confers Anti-Endothelial Adhesion and Anti-Migration Properties to Counteract Highly Metastatic Cervical Cancer Cells’ Migration in a Parallel-Plate Flow Chamber

Abstract

Despite the distant metastasis of cervical cancer cells being a prominent cause of mortality, neither the metastasis capacity nor the in vitro conditions mimicking adhesion of cervical cancer cells to endothelial cells have been fully elucidated. Circulating metastatic cancer cells undergo transendothelial migration and invade normal organs in distant metastasis; however, the putative molecular mechanism remains largely uncertain. In this study, we describe the use of an in vitro parallel-plate flow chamber to simulate the dynamic circulation stress on cervical cancer cells and elucidate their vascular adhesion and metastasis. We isolate the viable and shear stress-resistant (SSR) cervical cancer cells for mechanistic studies. Remarkably, the identified SSR-HeLa and SSR-CaSki exhibited high in vitro adhesive and metastatic activities. Hence, a consistently suppressed miR-128 level was revealed in SSR cell clones compared to those of parental wild-type (WT) cells. Overexpressed miR-128 attenuated SSR-HeLa cells’ adherence to human umbilical cord vein endothelial cells (HUVECs); in contrast, suppressed miR-128 efficiently augmented the static adhesion capacity in WT-HeLa and WT-CaSki cells. Hence, amplified miR-128 modestly abolished in vitro SSR-augmented HeLa and CaSki cell movement, whereas reduced miR-128 aggravated the migration speed in a time-lapse recording assay in WT groups. Consistently, the force expression of miR-128 alleviated the SSR-enhanced HeLa and CaSki cell mobility in a wound healing assay. Notably, miR-128 mediated SSR-enhanced HeLa and CaSki cells’ adhesion and metastasis through suppressed ITGA5, ITGB5, sLex, CEACAM-6, MMP9, and MMP23 transcript levels. Our data provide evidence suggesting that miR-128 is a promising microRNA that prevented endothelial cells’ adhesion and transendothelial migration to contribute to the SSR-enhanced adhesion and metastasis progression under a parallel-plate flow chamber system. This indicates that the nucleoid-based miR-128 strategy may be an attractive therapeutic strategy to eliminate tumor cells resistant to circulation shear flow, prevent vascular adhesion, and preclude subsequent transendothelial metastasis.

Adhesion to the Brain Endothelium Selects Breast Cancer Cells with Brain Metastasis Potential

“…Therefore, this study provides the first piece of evidence to demonstrate that the re-attachment to the brain endothelium enriches CTCs with brain metastatic potential. This is consistent with the previous finding that cervical cancer cells selected through the adhesion to the endothelium under fluid shear stress for 48 h exhibit a high metastatic potential [ 20 ]. The influence of this selection process may involve the potential effect of long-time exposure (48 h) to fluid shear stress, while the short-time selection (15 min) in this study mainly reflects the influence of tumor cell–endothelium adhesion on brain metastasis ability.…”

“…Tumor cells must adhere to the brain endothelium before extravasating into the tissue to generate brain metastases. To mimic the interaction between the endothelium and CTCs, we developed an in vitro system ( Figure 1 a) [ 19 , 20 , 21 ], in which a monolayer of brain endothelium (hCMEC/D3) was grown in the microfluidic channel and tumor cells in suspension were circulated under a steady flow in the system. Since CTCs are very rare in vivo (1–10 CTCs per mL blood) and not commercially available yet, tumor cells in suspension have been widely utilized as an alternative model for CTC study [ 22 , 23 , 24 , 25 , 26 ].…”

PIEZO1 Ion Channels Mediate Mechanotransduction in Odontoblasts

“…In contrast, we speculated that when external stimuli cause fluid flow into dentinal tubules, odontoblasts sense this FSS and convert it into electrical signals that are then transmitted to the nerve fibers to trigger a toothache. Based on the above hypothesis, we constructed an FSS model base on the previous literature 18,19 .…”

“…We established a mechanical stimulation model of fluid shear force to stimulate odontoblasts according to previous reports 18,19 . Fluo-4 AM was diluted with HBSS (with Ca 2+ and Mg 2+ ) to 2 mM and added to the OLC culture medium for 20 minutes.…”

Effect of Matrix Metalloproteinase 23 Accelerating Wound Healing Induced by Hydroxybutyl Chitosan

“…MMPs hydrolyze ECM components to influence their dynamic balance of degradation and restructuring, participate in a variety of cell physiological and pathological process, and influence wound healing and reconstruction. MMP23 is a unique component of the MMP protein family with characteristic domains and functions, degrading ECM proteins, cleaving cell–surface receptors, releasing apoptotic ligands, and activating chemokines and cytokines. − …”