Human Tumor Tissue-Based 3D In Vitro Invasion Assays

Åström, Pirjo; Heljasvaara, Ritva; Nyberg, Pia; Al‐Samadi, Ahmed; Salo, Tuula

doi:10.1007/978-1-4939-7595-2_19

Cited by 13 publications

(22 citation statements)

References 7 publications

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“…Cancer cell invasion was assessed in the myoma organotypic cultures and performed according to the previously published myoma model protocol 17,18 . Briefly, uterine leiomyoma tissues were obtained from routine surgery after informed consent of the donors.…”

Section: Myoma Organotypic Invasion Modelmentioning

confidence: 99%

New role of the antidepressant imipramine as a Fascin1 inhibitor in colorectal cancer cells

Alburquerque-González

Bernabé-García

Montoro-García

et al. 2020

Exp Mol Med

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Serrated adenocarcinoma (SAC) is more invasive, has worse outcomes than conventional colorectal carcinoma (CRC), and is characterized by frequent resistance to anti-epidermal growth factor receptor (EGFR) and overexpression of fascin1, a key protein in actin bundling that plays a causative role in tumor invasion and is overexpressed in different cancer types with poor prognosis. In silico screening of 9591 compounds, including 2037 approved by the Food and Drug Administration (FDA), was performed, and selected compounds were analyzed for their fascin1 binding affinity by differential scanning fluorescence. The results were compared with migrastatin as a typical fascin1 inhibitor. In silico screening and differential scanning fluorescence yielded the FDA-approved antidepressant imipramine as the most evident potential fascin1 blocker. Biophysical and different in vitro actin-bundling assays confirm this activity. Subsequent assays investigating lamellipodia formation and migration and invasion of colorectal cancer cells in vitro using 3D human tissue demonstrated anti-fascin1 and anti-invasive activities of imipramine. Furthermore, expression profiling suggests the activity of imipramine on the actin cytoskeleton. Moreover, in vivo studies using a zebrafish invasion model showed that imipramine is tolerated, its anti-invasive and antimetastatic activities are dose-dependent, and it is associated with both constitutive and induced fascin1 expression. This is the first study that demonstrates an antitumoral role of imipramine as a fascin1 inhibitor and constitutes a foundation for a molecular targeted therapy for SAC and other fascin1-overexpressing tumors.

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Section: Myoma Organotypic Invasion Modelmentioning

confidence: 99%

New role of the antidepressant imipramine as a Fascin1 inhibitor in colorectal cancer cells

Alburquerque-González

Bernabé-García

Montoro-García

et al. 2020

Exp Mol Med

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“…The complexity of TME is, after all, beyond any in vitro models, as they lack the elements that angiogenesis affects (vascularity) as well as inflammatory cells and other possible proteins or co-factors that may be involved in the pathways of enzyme activity. However, we wanted to mimic the TME in our in vitro experiments, and hence, also used our human myoma-derived organotypic model to more reliably evaluate the effects of MMP9 on cell behaviour [22,23]. Because of the diversity of the effects and functions of MMP9 reported in vitro, it would be highly important to evaluate whether the upregulated MMP9 in vivo has a true pathogenic effect or whether the upregulation is actually caused by the disease.…”

Section: Discussionmentioning

confidence: 99%

“…Myoma organotypic cultures were prepared as previously described [22,23]. Briefly, uterine leiomyoma tissue was obtained during routine surgeries with the informed consent of the donors.…”

Section: Myoma Organotypic Cultures and Immunohistochemistrymentioning

confidence: 99%

“…The tissues were fixed in 4% neutral-buffered formalin overnight, dehydrated and embedded in paraffin. Then 6-μm sections were deparaffinized and endogenous peroxidase was blocked with H 2 O 2 in methanol as the specimens were prepared for immunohistochemistry with monoclonal pan-cytokeratin AE1/AE3 antibody (Dako) to identify carcinoma cells, and invasion was quantified as described earlier [22,23]. Briefly, histological sections were photographed with a DMRB photo microscope connected to a DFC-480 camera using QWin V3 software (Leica Microsystems) or with an Olympus BX61 light-field microscope equipped with an Olympus U-CMAB3 camera.…”

Section: Myoma Organotypic Cultures and Immunohistochemistrymentioning

confidence: 99%

“…Briefly, histological sections were photographed with a DMRB photo microscope connected to a DFC-480 camera using QWin V3 software (Leica Microsystems) or with an Olympus BX61 light-field microscope equipped with an Olympus U-CMAB3 camera. The invasion depth and areas of non-invading and invading cells were measured with QWin V3 software, and the invasion index was calculated as previously described [22,23].…”

Section: Myoma Organotypic Cultures and Immunohistochemistrymentioning

confidence: 99%

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Matrix metalloproteinase 9 inhibits the motility of highly aggressive HSC-3 oral squamous cell carcinoma cells

Väyrynen

Åström

Nyberg

et al. 2019

Experimental Cell Research

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Pro-tumorigenic activities of matrix metalloproteinase (MMP) 9 have been linked to many cancers, but recently the tumour-suppressing role of MMP9 has also been elucidated. The multifaceted evidence on this subject prompted us to examine the role of MMP9 in the behaviour of oral tongue squamous cell carcinoma (OTSCC) cells. We used gelatinase-specific inhibitor, CTT2, and short hairpin (sh) RNA gene silencing to study the effects of MMP9 on proliferation, motility and invasion of an aggressive OTSCC cell line, HSC-3. We found that the migration and invasion of HSC-3 cells were increased by CTT2 and shRNA silencing of MMP9. Proliferation, in turn, was decreased by MMP9 inhibition. Furthermore, arresten-overexpressing HSC-3 cells expressed increased levels of MMP9, but exhibited decreased motility compared with controls. Interestingly, these cells restored their migratory capabilities by CTT2 inhibition of MMP9. Hence, although higher MMP9 expression could give rise to an increased tumour growth in vivo due to increased proliferation, in some circumstances, it may participate in yet unidentified molecular mechanisms that reduce the cell movement in OTSCC.

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SLC4A2 anion exchanger promotes tumour cell malignancy via enhancing net acid efflux across golgi membranes

Khosrowabadi

Rivinoja²,

Risteli

et al. 2021

Cell. Mol. Life Sci.

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Proper functioning of each secretory and endocytic compartment relies on its unique pH micro-environment that is known to be dictated by the rates of V-ATPase-mediated H+ pumping and its leakage back to the cytoplasm via an elusive “H+ leak” pathway. Here, we show that this proton leak across Golgi membranes is mediated by the AE2a (SLC4A2a)-mediated bicarbonate-chloride exchange, as it is strictly dependent on bicarbonate import (in exchange for chloride export) and the expression level of the Golgi-localized AE2a anion exchanger. In the acidic Golgi lumen, imported bicarbonate anions and protons then facilitate a common buffering reaction that yields carbon dioxide and water before their egress back to the cytoplasm via diffusion or water channels. The flattened morphology of the Golgi cisternae helps this process, as their high surface-volume ratio is optimal for water and gas exchange. Interestingly, this net acid efflux pathway is often upregulated in cancers and established cancer cell lines, and responsible for their markedly elevated Golgi resting pH and attenuated glycosylation potential. Accordingly, AE2 knockdown in SW-48 colorectal cancer cells was able to restore these two phenomena, and at the same time, reverse their invasive and anchorage-independent growth phenotype. These findings suggest a possibility to return malignant cells to a benign state by restoring Golgi resting pH.

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Human Tumor Tissue-Based 3D In Vitro Invasion Assays

Cited by 13 publications

References 7 publications

New role of the antidepressant imipramine as a Fascin1 inhibitor in colorectal cancer cells

New role of the antidepressant imipramine as a Fascin1 inhibitor in colorectal cancer cells

Matrix metalloproteinase 9 inhibits the motility of highly aggressive HSC-3 oral squamous cell carcinoma cells

SLC4A2 anion exchanger promotes tumour cell malignancy via enhancing net acid efflux across golgi membranes

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