The autophagy inhibitor spautin-1, either alone or combined with doxorubicin, decreases cell survival and colony formation in canine appendicular osteosarcoma cells

Schott, Courtney R.; Ludwig, Latasha; Mutsaers, Anthony J.; Foster, Robert A.; Wood, Geoffrey A.

doi:10.1371/journal.pone.0206427

Cited by 31 publications

(25 citation statements)

References 72 publications

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“…In this study, effort was invested on optimizing a specific assay through use of multiple antibodies to exclude nonspecific antibody binding to extracellular or intracellular epitopes of OSA cells, distinction of leukocytes from OSA cells, and assuring that only viable single cells were being assessed. Initial optimization experiments utilizing cell lines derived from a primary and a metastatic tumor showed slightly differing light scatter properties, consistent also with differing biological properties and signaling pathways of these cell lines (33). Both cell lines had moderate to high FSC but OVC-cOSA-75 cells had low SSC.…”

Section: Discussionmentioning

confidence: 75%

“…Cell lines OVC-cOSA-75 and OVC-cOSA-31 were derived from autopsy and amputation specimens from two separate dogs with lung metastases or primary OSA of the distal tibia, respectively (33). Cell lines were established and maintained as monolayers in DMEM (Sigma Aldrich, Oakville, ON) supplemented with 10% fetal bovine serum (Fisher Scientific, Ottawa, ON) and 1% penicillin/streptomycin (Sigma Aldrich), and cultured at 37 C in a humidified incubator in the presence of 95% atmospheric air and 5% CO 2 .…”

Section: Immortalized Osa Cell Linesmentioning

confidence: 99%

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Flow Cytometric Detection of Circulating Osteosarcoma Cells in Dogs

et al. 2019

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Osteosarcoma (OSA) is a malignant tumor of middle-aged dogs and adolescent humans. The clinical outcome of OSA has not improved over more than three decades, and dogs typically succumb to metastatic disease within 6 months despite tumor resection through limb amputation and adjuvant chemotherapy. Therefore, undetectable tumor cells with potential to form metastases are present at diagnosis. An assay to identify canine immortalized and primary OSA cells through flow cytometric detection of intracellular collagen 1 (Col I) and osteocalcin was optimized, and applied to blood samples from tumorbearing dogs for detection of circulating tumor cells (CTCs). Spiking variable number of OSA cells into normal dog blood recovered 50-60% of Col I positive cells with high forward and variable side light scatter. An algorithm to exclude nonviable, doublet, and autofluorescent cells was applied to sequential blood samples from three dogs obtained prior to and after limb amputation, and at approximately, triweekly intervals over 121, 142, and 183 days of chemotherapy, respectively. Dogs had >100 CTC/10 6 leukocytes prior to amputation, variably frequent CTC during chemotherapy, and an increase up to 4,000 CTC/10 6 leukocytes within 4 weeks before overt metastases or death. Sorted CTCs were morphologically similar to direct tumor aspirates and positive for Col I. Although preliminary, findings suggest that CTCs are frequent in canine OSA, more numerous than carcinoma CTC in humans, and that an increase in CTC frequency may herald clinical deterioration. This assay may enable enumeration and isolation of OSA CTC for prognostic and functional studies, respectively. OSTEOSARCOMA (OSA) is a primary bone cancer that is common in middle-aged to older, large breed dogs, and less common in humans, where children and young adults are most often affected. Therapy consisting of limb amputation or limbsparing tumor resection followed by chemotherapy (typically platinum or doxorubicin-based) is the gold standard, but the clinical outcome of OSA in dogs and humans remains poor (1-3). Dogs with OSA of the appendicular skeleton treated with amputation and chemotherapy had median survival times (STs) of 235-540 days (4), while in humans 5-year survival has been stagnant at about 60% (5). Although the median ST is relatively homogeneous, the range of ST in dogs with OSA treated identically varies from 2 to >30 months, illustrating that there is extensive heterogeneity in tumor biology and/or host response (6-8). Metastatic disease is the usual cause of spontaneous death or euthanasia, but fewer than 10% of dogs have radiographic evidence of metastasis at the time of diagnosis (2).Computed tomography also lacks sensitivity for identifying metastases, and histologic grading schemes for canine OSA poorly predict biological behavior (9-11).

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Section: Discussionmentioning

confidence: 75%

Section: Immortalized Osa Cell Linesmentioning

confidence: 99%

Flow Cytometric Detection of Circulating Osteosarcoma Cells in Dogs

et al. 2019

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“…The chemotherapeutic agent Dox induces autophagy in human osteosarcoma cell lines [23]. Autophagy contributes to chemoresistance in canine appendicular osteosarcoma and adding an autophagy inhibitor to the standard of care has the potential to improve outcome [15]. In the present study, we constructed a Dox-resistant U2OS cell subtype, U2OS/Dox, and consistently observed that autophagy inhibitor 3-MA could resensitize U2OS/Dox to Dox treatment.…”

Section: Agingmentioning

confidence: 56%

“…Autophagy might be one mechanism by which neoplastic cells maintain survival upon chemotherapy treatment. Under such conditions, blocking autophagy could trigger apoptosis, therefore enhancing the curative effects of these chemotherapies and reducing chemoresistance [15,16]. Here, we constructed a Doxresistant U2OS cell line, U2OS/Dox, by treating…”

Section: Dox Treatment Induces Autophagy and Sox2ot-v7 Expression In mentioning

confidence: 99%

LncRNA Sox2OT-V7 promotes doxorubicin-induced autophagy and chemoresistance in osteosarcoma via tumor-suppressive miR-142/miR-22

Zhu

Yang

Wen

et al. 2020

Aging

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Doxorubicin (Dox) is one of the most commonly used chemotherapeutic drugs for osteosarcoma (OS) treatment. In the present study, we attempted to investigate the mechanism by which Sox2OT-V7 dysregulation affects Dox chemoresistance to provide a novel experimental basis for developing neoadjuvant therapy. Sox2OT-V7 expression is upregulated in OS tissues, particularly in chemoresistant OS tissues, and in OS cell lines compared to controls. Dox treatment induces autophagy and Sox2OT-V7 expression in U2OS cells, and Dox-induced autophagy is partially attenuated by Sox2OT-V7 silencing. Knocking down Sox2OT-V7 or blocking autophagy in Dox-resistant U2OS/Dox cells resensitizes the cells to Dox treatment in vitro. Moreover, Sox2OT-V7 directly targets miR-142/miR-22 to inhibit their expression, and the effect of Sox2OT-V7 silencing on U2OS cell autophagy and U2OS/Dox cell sensitivity to Dox can be reversed by miR-142/miR-22 inhibition. Sox2OT-V7 silencing enhances the suppressive effects of Dox on U2OS/Dox cell-derived tumor growth in vivo, while miR-22 inhibition or miR-142 inhibition reverses the effects of Sox2OT-V7 silencing on Dox-induced suppression on tumor growth. Finally, miR-142 directly targets ULK1, ATG4A, and ATG5, while miR-22 directly targets ULK1 to inhibit the expression of the target gene; The Sox2OT-V7/miR-142/miR-22 axis modulates autophagy in OS cells by regulating ULK1, ATG4A, and ATG5.

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“…Considering the role of autophagy in OS treatment resistance, many studies combined the use of anti-cancer agents with autophagy inhibitors such as chloroquine, 3-methyladenine or spautin-1 (specific and potent autophagy inhibitor-1). These autophagy inhibitors enhance OS cell death induced by cisplatin [73], doxorubicin [74], bone-targeted gallium compound KP46 [75], and increase in vivo tumor regression following photodynamic therapy [76] or mTOR inhibition [77].…”

Section: Autophagy In Osteosarcomamentioning

confidence: 99%

Role of autophagy in osteosarcoma

Camuzard

Santucci‐Darmanin

Carle

et al. 2019

Journal of Bone Oncology

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Osteosarcoma (OS) is the most common primary bone tumour in children and adolescents. It is a highly aggressive tumor with a tendency to spread to the lungs, which are the most common site of metastasis. Advanced osteosarcoma patients with metastasis share a poor prognosis. Despite the use of chemotherapy to treat OS, the 5-year overall survival rate for patients has remained unchanged at 65–70% for the past 20 years. In addition, the 5-year survival of patients with a metastatic disease is around 20%, highlighting the need for novel therapeutic targets. Autophagy is an intracellular degradation process which eliminates and recycles damaged proteins and organelles to improve cell lifespan. In the context of cancer, numerous studies have demonstrated that autophagy is used by tumor cells to repress initial steps of carcinogenesis and/or support the survival and growth of established tumors. In osteosarcoma, autophagy appears to be deregulated and could also act both as a pro or anti-tumoral process. In this manuscript, we aim to review these major findings regarding the role of autophagy in osteosarcoma.

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The autophagy inhibitor spautin-1, either alone or combined with doxorubicin, decreases cell survival and colony formation in canine appendicular osteosarcoma cells

Cited by 31 publications

References 72 publications

Flow Cytometric Detection of Circulating Osteosarcoma Cells in Dogs

Flow Cytometric Detection of Circulating Osteosarcoma Cells in Dogs

LncRNA Sox2OT-V7 promotes doxorubicin-induced autophagy and chemoresistance in osteosarcoma via tumor-suppressive miR-142/miR-22

Role of autophagy in osteosarcoma

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