Primary Tumor and MEF Cell Isolation to Study Lung Metastasis

Dong, Shaohong; Maziveyi, Mazvita

doi:10.3791/52609

Cited by 9 publications

(6 citation statements)

References 12 publications

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“…For mouse genotyping, mouse tail genomic DNA was extracted and amplified by PCR and electrophoresed on 2% agarose gels. Primary WT-PyMT (Nisch þ/þ ), HET-PyMT (Nisch þ/À ), and Null-PyMT (Nisch À/À ) cells were isolated as described previously (17). Briefly, the mammary tumors were isolated and cut into small pieces with a razor blade and scissors.…”

Section: Cell Culturementioning

confidence: 99%

“…The supernatant was then discarded and the pellet was centrifuged with PBS for 70 minutes at 1 Â 100,000 Â g. The pellet was finally resuspended in PBS for subsequent studies. To isolate exosomes from whole tumors, we harvested breast tumors from Nisch þ/þ and Nisch þ/À mice as described previously (17). Tumors were disintegrated and incubated with collagenase for 3 hours.…”

Section: Isolation Of Exosomesmentioning

confidence: 99%

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Exosomes from Nischarin-Expressing Cells Reduce Breast Cancer Cell Motility and Tumor Growth

et al. 2019

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Exosomes are small extracellular microvesicles that are secreted by cells when intracellular multivesicular bodies fuse with the plasma membrane. We have previously demonstrated that Nischarin inhibits focal adhesion formation, cell migration, and invasion, leading to reduced activation of focal adhesion kinase. In this study, we propose that the tumor suppressor Nischarin regulates the release of exosomes. When cocultured on exosomes from Nischarin-positive cells, breast cancer cells exhibited reduced survival, migration, adhesion, and spreading. The same cocultures formed xenograft tumors of significantly reduced volume following injection into mice. Exosomes secreted by Nischarin-expressing tumors inhibited tumor growth. Expression of only one allele of Nischarin increased secretion of exosomes, and Rab14 activity modulated exosome secretions and cell growth. Taken together, this study reveals a novel role for Nischarin in preventing cancer cell motility, which contributes to our understanding of exosome biology. Significance: Regulation of Nischarin-mediated exosome secretion by Rab14 seems to play an important role in controlling tumor growth and migration. See related commentary by McAndrews and Kalluri, p. 2099

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Section: Cell Culturementioning

confidence: 99%

Section: Isolation Of Exosomesmentioning

confidence: 99%

Exosomes from Nischarin-Expressing Cells Reduce Breast Cancer Cell Motility and Tumor Growth

et al. 2019

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“…The lungs were excised, filled with dilute India ink solution through the trachea, destained in Fatake’s solution, and fixed in 4% paraformaldehyde. 32–34 The number of metastatic nodules were counted.…”

Section: Methodsmentioning

confidence: 99%

“…On day 33 postinoculation, mice were sacrificed, and primary tumors in the mammary fat pad were excised and weighed. The lungs were excised, filled with dilute India ink solution through the trachea, destained in Fatake’s solution, and fixed in 4% paraformaldehyde. − The number of metastatic nodules was counted.…”

Section: Methodsmentioning

confidence: 99%

Magnetophoretic Delivery of a Tumor-Priming Agent for Chemotherapy of Metastatic Murine Breast Cancer

Park

Castanares

et al. 2019

Mol. Pharmaceutics

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Tumor microenvironment (TME) is a significant physical barrier to effective delivery of chemotherapy into solid tumors. To overcome this challenge, tumors are pre-treated with an agent that reduces cellular and extracellular matrix densities prior to chemotherapy. However, it also comes with a concern that metastasis may increase due to the loss of protective containment. We hypothesize that timely priming at the early stage of primary tumors will help control metastasis. To test this, we primed orthotopic 4T1 breast tumors with a paclitaxel (PTX)-loaded iron oxide decorated poly(lactic-co-glycolic acid) nanoparticle composite (PTX@PINC), which can be quickly concentrated in target tissues with the aid of an external magnet, and monitored its effect on the delivery of subsequently administered NPs. Magnetic resonance imaging and optical whole-body imaging confirmed that PTX@PINC was efficiently delivered to tumors by the external magnet and help loosen the tumors to accommodate subsequently-delivered NPs. Consistently, the primed tumors responded to Doxil better than non-primed tumors. In addition, lung metastasis was significantly reduced in the animals PINC-primed prior to Doxil administration. These results support that PINC combined with magnetophoresis can facilitate timely management of primary tumors with a favorable secondary effect on metastasis.

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“…Mouse embryonic fibroblasts (MEFs): E13.5 embryos were isolated from timed matings of C57Bl/6J mice. Internal organs and heads were removed and tissue homogenized by mincing with razor blades and pressure from the back of a syringe (72)(73)(74). Single cell suspensions were further generated by a 30-min incubation at 37 C in 5 ml Trypsin-EDTA.…”

Section: Cellsmentioning

confidence: 99%

Cell-Intrinsic and Extrinsic Effects of Galectin-1 and Galectin-3 in B-Cell Precursor Acute Lymphoblastic Leukemia

Fei

Joo

Yang

et al. 2021

Preprint

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Acute lymphoblastic leukemias arising from the malignant transformation of B-cell precursors (BCP ALLs) are protected against chemotherapy by both intrinsic factors as well as by interactions with bone marrow stromal cells. Galectin-1 and Galectin-3 have overlapping expression patterns and potentially common functions in these cells. We used Galectin-1 and Galectin-3 double null mutant murine BCP ALL cells to examine the effect of loss endogenous Galectins. We also tested the effect of Galectin inhibition by use of plant-derived natural compounds GM-CT-01 and GR-MD-02 in human BCP ALL cells in co-culture with stroma. Transformed wild type and Galectin-1 x Galectin-3 double knockout BCP ALL cells were similar in immunophenotype and active intracellular signaling. However, compared to wild type cells, they showed impaired migration, significantly reduced proliferation and increased sensitivity to drug treatment. GM-CT-01 and GR-MD-02 attenuated intracellular signal transduction and sensitized the ALL cells to chemotherapy including vincristine and the targeted tyrosine kinase inhibitor nilotinib. Our data show endogenous and extracellular Galectins contribute positively to the growth and survival of BCP ALL cells. Strategies that would efficiently ablate both Galectins at both locations would decrease microenvironmental protection and reduce BCP ALL persistence in the protective bone marrow niche after chemotherapy.

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Primary Tumor and MEF Cell Isolation to Study Lung Metastasis

Cited by 9 publications

References 12 publications

Exosomes from Nischarin-Expressing Cells Reduce Breast Cancer Cell Motility and Tumor Growth

Exosomes from Nischarin-Expressing Cells Reduce Breast Cancer Cell Motility and Tumor Growth

Magnetophoretic Delivery of a Tumor-Priming Agent for Chemotherapy of Metastatic Murine Breast Cancer

Cell-Intrinsic and Extrinsic Effects of Galectin-1 and Galectin-3 in B-Cell Precursor Acute Lymphoblastic Leukemia

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