Antibiotics inhibit sphere-forming ability in suspension culture

Relier, Sébastien; Yazdani, Laura; Ayad, Oualid; Choquet, Armelle; Bourgaux, Jean–François; Prudhomme, Michel; Pannequin, Julie; Macari, Françoise; David, Alexandre

doi:10.1186/s12935-016-0277-6

Cited by 20 publications

(22 citation statements)

References 27 publications

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“…However, this may not apply to different cancers or even other, not assayed, RCC cell lines (41). Penicillin-streptomycin enhanced Caki-1 2D cell growth as observed previously for colon cancer cell lines (42), but its mechanism requires further analysis. The tested RCC cell lines were also insensitive to azithromycin, both the 2D and the sphere-forming population.…”

Section: Discussionsupporting

confidence: 65%

“…It is also worth emphasizing that sunitinib suppresses the proliferation, migration, apoptosis resistance, tumor angiogenesis and growth of triple-negative breast cancers but increases the percentage of breast cancer stem cells in tumors (39), which is in agreement with the fact that sunitinib was recently shown in preclinical models to increase invasive and metastatic properties of breast cancer cells (40). Surprisingly, recently, several groups have reported that antibiotics, including those routinely used in cell culture, can interfere with the sphere-forming abilities of cancer cells, and therefore their impact on CSCs may be expected (41,42). Selective toxic effects on CSCs have been reported for azithromycin, chloramphenicol and doxycycline (41).…”

Section: Introductionmentioning

confidence: 99%

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Metastatic renal cell carcinoma cells growing in 3D on poly‑D‑lysine or laminin present a stem‑like phenotype and drug resistance

Brodaczewska¹,

Bielecka²,

Maliszewska‐Olejniczak³

et al. 2019

Oncol Rep

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3D spheroids are built by heterogeneous cell types in different proliferative and metabolic states and are enriched in cancer stem cells. The main aim of the study was to investigate the usefulness of a novel metastatic renal cell carcinoma (RCC) 3D spheroid culture for in vitro cancer stem cell physiology research and drug toxicity screening. RCC cell lines, Caki-1 (skin metastasis derived) and ACHN (pleural effusion derived), were efficiently cultured in growth-factor/serum deprived, defined, StemXvivo and Nutristem medium on laminin-coated or poly-D-lysine-coated plates. In optimal 3D culture conditions, ACHN cells (StemXVivo/poly-D-lysine) formed small spheroids with remaining adherent cells of an epithelial phenotype, while Caki-1 cells (StemXVivo/laminin) formed large dark spheroids with significantly reduced cell viability in the center. In the 3D structures, expression levels of genes encoding stem transcription factors (OCT4, SOX2, NES) and RCC stem cell markers (CD105, CD133) were deregulated in comparison to these expression levels in traditional 2D culture. Sunitinib, epirubicin and doxycycline were more toxic to cells cultured in monolayers than for cells in 3D spheroids. High numbers of cells arrested in the G0/G1 phase of the cell cycle were found in spheroids under sunitinib treatment. We showed that metastatic RCC 3D spheroids supported with ECM are a useful model to determine the cancer cell growth characteristics that are not found in adherent 2D cultures. Due to the more complex architecture, spheroids may mimic in vivo micrometastases and may be more appropriate to investigate novel drug candidate responses, including the direct effects of tyrosine kinase inhibitor activity against RCC cells.

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Section: Discussionsupporting

confidence: 65%

Section: Introductionmentioning

confidence: 99%

Metastatic renal cell carcinoma cells growing in 3D on poly‑D‑lysine or laminin present a stem‑like phenotype and drug resistance

Brodaczewska¹,

Bielecka²,

Maliszewska‐Olejniczak³

et al. 2019

Oncol Rep

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“…However, it also possesses some affinity to the ribosomes in eukaryotic cells. Relier and colleagues suggested that streptomycin may act on eukaryotic cells through the disruption of protein synthesis, by affecting mitochondrial activity, by binding to certain RNAs or by interfering with miRNAs action [ 12 ]. This indicates that the addition of streptomycin changes the gene expression profile in cells and it may affect the differentiation process.…”

Section: Discussionmentioning

confidence: 99%

“…Isolation and cultivation of adipose-derived stem cells usually involves the presence of the penicillin-streptomycin mixture [ 2 , 5 , 8 , 9 ] although some studies have also suggested using amphotericin B because of its widespread antifungal activity [ 10 , 11 ]. Gentamycin is also commonly used in cell cultures, although a penicillin-streptomycin (PS) mixture is the only one that is recommended by the American Type Culture Collection (ATCC) [ 12 ]. Protective/preventive antibiotic therapy improves the success/safety of transplantation procedures [ 10 ].…”

Section: Introductionmentioning

confidence: 99%

Impact of Antibiotics on the Proliferation and Differentiation of Human Adipose-Derived Mesenchymal Stem Cells

Skubis

Gola

Sikora

et al. 2017

IJMS

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Adipose tissue is a promising source of mesenchymal stem cells. Their potential to differentiate and regenerate other types of tissues may be affected by several factors. This may be due to in vitro cell-culture conditions, especially the supplementation with antibiotics. The aim of our study was to evaluate the effects of a penicillin-streptomycin mixture (PS), amphotericin B (AmB), a complex of AmB with copper (II) ions (AmB-Cu2+) and various combinations of these antibiotics on the proliferation and differentiation of adipose-derived stem cells in vitro. Normal human adipose-derived stem cells (ADSC, Lonza) were routinely maintained in a Dulbecco’s Modified Eagle Medium (DMEM) that was either supplemented with selected antibiotics or without antibiotics. The ADSC that were used for the experiment were at the second passage. The effect of antibiotics on proliferation was analyzed using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) and sulforhodamine-B (SRB) tests. Differentiation was evaluated based on Alizarin Red staining, Oil Red O staining and determination of the expression of ADSC, osteoblast and adipocyte markers by real-time RT-qPCR. The obtained results indicate that the influence of antibiotics on adipose-derived stem cells depends on the duration of exposure and on the combination of applied compounds. We show that antibiotics alter the proliferation of cells and also promote natural osteogenesis, and adipogenesis, and that this effect is also noticeable in stimulated osteogenesis.

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“…This test was performed as previously described [61]. Number of sphere forming cells were determined after plating of 100 cells / 100 µL of M11 medium (DMEM/F12 (1:1) Glutamax medium, N2 Supplement, Glucose 0.3 %, insulin 20 µg/ml, hBasic-FGF 10 ng/ml, hEGF 20 ng/ml) in ultra-low attachment 96 well-plates.…”

Section: Methodsmentioning

confidence: 99%

FTO-mediated cytoplasmic m⁶A_mdemethylation adjusts stem-like properties in colorectal cancer cell

Relier

Ripoll

Guillorit

et al. 2020

Preprint

Self Cite

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20Cancer stem cells (CSCs) are a small but critical cell population for cancer biology since they display 21 inherent resistance to standard therapies and give rise to metastases. Despite accruing evidence 22 establishing a link between deregulation of epitranscriptome-related players and tumorigenic 23 process, the role of messenger RNA (mRNA) modifications dynamic in the regulation of CSC 24 properties remains poorly understood. Here, we show that the cytoplasmic pool of fat mass and 25 obesity-associated protein (FTO) impedes CSC abilities in colorectal cancer through its m 6 Am (N 6 ,2'-26 O-dimethyladenosine) demethylase activity. While m 6 Am is strategically located next to the m 7 G-27 mRNA cap, its biological function is not well understood and has not been addressed in cancer. Low 28 FTO expression in patient-derived cell lines elevates m 6 Am level in mRNA which results in enhanced 29 in vivo tumorigenicity and chemoresistance. Inhibition of the nuclear m 6 Am methyltransferase, 30 PCIF1/CAPAM, partially reverses this phenotype. FTO-mediated regulation of m 6 Am marking 31 constitutes a novel, reversible pathway controlling CSC abilities that does not involve transcriptome 32 remodeling, but could fine-tune translation efficiency of selected m 6 Am marked transcripts. 33 Altogether, our findings bring to light the first biological function of the m 6 Am modification and its 34 potential adverse consequences for colorectal cancer management. 35 36 Despite significant advances in diagnosis and therapy, colorectal cancer (CRC) remains a major 37 cause of mortality and morbidity worldwide. CRC survival is highly dependent upon early diagnosis. 38 Patients with localized cancer exhibit 70 to 90% 5-year survival. Survival from metastatic cancer 39 plummets to 10%. Metastasis is a multistep process encompassing local infiltration of tumor cells into 40 adjacent tissues, transendothelial migration into vessels, survival in the circulatory system, 41 extravasation, and colonization of secondary organs [1]. This process entails constant reprogramming 42 of gene expression to enable tumor adaptation in different environments, a peculiar trait of cancer 43 stem cells (CSCs). CSC constitute a minor subpopulation of tumor cells endowed with self-renewal and 44 multi-lineage differentiation capacity [2]. The most clinically relevant trait of CSCs is their ability to 45 metastasize and escape from standard chemotherapy [3]. Understanding the molecular mechanisms 46 that participate to the CSC phenotype is critical to designing improved cancer therapeutics. 47 Discovered several decades ago [12-16], the function of m 6 A remained obscure until the identification 55 of the first m 6 A demethylase, the fat mass and obesity-associated protein (FTO) [17]. The marriage of 56 immunochemical approaches with next-generation sequencing (NGS) technologies revealed the 57 unique topology of m 6 A distribution along mRNA. m 6 A is a dynamic reversible chemical modification 58 catalyzed by a protein complex consisting of the methyl...

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Antibiotics inhibit sphere-forming ability in suspension culture

Cited by 20 publications

References 27 publications

Metastatic renal cell carcinoma cells growing in 3D on poly‑D‑lysine or laminin present a stem‑like phenotype and drug resistance

Metastatic renal cell carcinoma cells growing in 3D on poly‑D‑lysine or laminin present a stem‑like phenotype and drug resistance

Impact of Antibiotics on the Proliferation and Differentiation of Human Adipose-Derived Mesenchymal Stem Cells

FTO-mediated cytoplasmic m⁶A_mdemethylation adjusts stem-like properties in colorectal cancer cell

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Antibiotics inhibit sphere-forming ability in suspension culture

Cited by 20 publications

References 27 publications

Metastatic renal cell carcinoma cells growing in 3D on poly‑D‑lysine or laminin present a stem‑like phenotype and drug resistance

Metastatic renal cell carcinoma cells growing in 3D on poly‑D‑lysine or laminin present a stem‑like phenotype and drug resistance

Impact of Antibiotics on the Proliferation and Differentiation of Human Adipose-Derived Mesenchymal Stem Cells

FTO-mediated cytoplasmic m6Amdemethylation adjusts stem-like properties in colorectal cancer cell

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FTO-mediated cytoplasmic m⁶A_mdemethylation adjusts stem-like properties in colorectal cancer cell