scholarly journals Effects of novel somatostatin-dopamine chimeric drugs in 2D and 3D cell culture models of neuroendocrine tumors

2019 ◽  
Vol 26 (6) ◽  
pp. 585-599 ◽  
Author(s):  
Aura D Herrera-Martínez ◽  
Rosanna van den Dungen ◽  
Fadime Dogan-Oruc ◽  
Peter M van Koetsveld ◽  
Michael D Culler ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Cell Growth ◽  
Dopamine Receptor ◽  
Neuroendocrine Tumors ◽  
3D Model ◽  
Serotonin Release ◽  
Receptor Expression ◽  
3D Cultures ◽  
2D And 3D ◽  
3D Spheroids

Control of symptoms related to hormonal hypersecretion by functioning neuroendocrine tumors (NETs) is challenging. New therapeutic options are required. Since novel in vitro tumor models seem to better mimic the tumor in vivo conditions, we aimed to study the effect of somatostatin and dopamine receptor agonists (octreotide and cabergoline, respectively) and novel somatostatin-dopamine chimeric multi-receptor drugs (BIM-065, BIM-23A760) using 2D (monolayer) and 3D (spheroids) cultures. Dose–response studies in 2D and 3D human pancreatic NET cell cultures (BON-1 and QGP-1) were performed under serum-containing and serum-deprived conditions. Cell proliferation, somatostatin and dopamine receptor expression (SSTs and D2R), apoptosis, lactate dehydrogenase, as well as serotonin and chromogranin A (CgA) release were assessed. The following results were obtained. 3D cultures of BON-1/QGP-1 allowed better cell survival than 2D cultures in serum-deprived conditions. SSTs and D2R mRNA levels were higher in the 3D model vs 2D model. Octreotide/cabergoline/BIM-065/BIM-23A760 treatment did not affect cell growth or spheroid size. In BON-1 2D-cultures, only BIM-23A760 significantly inhibited CgA release –this effect being more pronounced in 3D cultures. In BON-1 2D cultures, cabergoline/BIM-065/BIM-23A760 treatment decreased serotonin release (maximal effect up to 40%), being this effect again more potent in 3D cultures (up to 67% inhibition; with BIM-23A760 having the most potent effects). In QGP-1, cabergoline/BIM-065 treatment decreased serotonin release only in the 3D model. In conclusion, cultures of NET 3D spheroids represent a promising method for evaluating cell proliferation and secretion in NET cell-line models. Compared to 2D models, 3D models grow relatively serum independent. In 3D model, SST-D2R multi-receptor targeting drugs inhibit CgA and serotonin secretion, but not NET cell growth.

scholarly journals Surface Acoustic Waves (SAW)-Based Biosensing for Quantification of Cell Growth in 2D and 3D Cultures

Sensors ◽  
2015 ◽  
Vol 15 (12) ◽  
pp. 32045-32055 ◽  
Author(s):  
Tao Wang ◽  
Ryan Green ◽  
Rajesh Nair ◽  
Mark Howell ◽  
Subhra Mohapatra ◽  
...  
Keyword(s):  
Cell Growth ◽  
Acoustic Waves ◽  
Surface Acoustic Waves ◽  
3D Cultures ◽  
2D And 3D

Somatostatin and dopamine receptor expression in lung carcinoma cells and effects of chimeric somatostatin-dopamine molecules on cell proliferation

2005 ◽  
Vol 289 (6) ◽  
pp. E1044-E1050 ◽  
Author(s):  
Diego Ferone ◽  
Marica Arvigo ◽  
Claudia Semino ◽  
Philippe Jaquet ◽  
Alexandru Saveanu ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Cell Growth ◽  
Inhibitory Effect ◽  
Cell System ◽  
Receptor Expression ◽  
Suitable Model ◽  
Independent Manner ◽  
Lung Carcinoma Cells ◽  
Cancer Line ◽  
Direct Inhibitory Effect

To study somatostatin/dopamine (SS/D) synergy in a human cell system constitutively expressing SS and D receptors (SSR and DR, respectively), we characterized the expression of SSR and DR subtypes in the non-small-cell lung cancer line Calu-6, and then we evaluated the effect on cell proliferation of SS/D chimeric molecules (BIM-23A387 and BIM-23A370), which bind with high affinity both sst2 and D2R, and compared the results with those obtained by using SS-14 and subtype-selective SS analogs (SSA) and D agonists (DA). Because Calu-6 cells produce insulin-like growth factor (IGF) and IGF-binding protein (IGFBP) peptides, which play a role in the autocrine/paracrine control of cell growth, we also investigated the effects of chimeric compounds on secretion and expression of IGF system components. Relative high levels of sst2 and the long isoform of the D2R were detected by real-time RT-PCR and Western blot in Calu-6, together with sst5 and to a lesser extent sst3 and D4R. BIM-23A387 and BIM-23A370 significantly inhibited growth of Calu-6, whereas IGF-IGFBP secretion or expression was unaffected, suggesting a direct inhibitory effect. The inhibition of cell growth, measured by both [3H]thymidine incorporation and cell count, was significantly lower when individual SSA and DA control peptides or subtype-specific SSA and DA were tested. BIM-23A370 was more potent than BIM-23A387 ( P < 0.001). These findings show that SS/D chimeras can inhibit Calu-6 proliferation in an IGF-independent manner and suggest that this enhanced potency might be because of the induction of SSR/DR dimerization. The Calu-6 cell line, constitutively expressing SSR and DR, provides a suitable model to elucidate the mechanism of action of SSA and DA on regulation of cell growth and to characterize the interaction between SSR and DR.

scholarly journals Three dimensional cultivation increases chemo- and radioresistance of colorectal cancer cell lines

PLoS ONE ◽  
2021 ◽  
Vol 16 (1) ◽  
pp. e0244513
Author(s):  
Jana Koch ◽  
Dina Mönch ◽  
Annika Maaß ◽  
Christian Gromoll ◽  
Thomas Hehr ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Cell Lines ◽  
Cell Cultures ◽  
Therapy Response ◽  
Fold Increase ◽  
3D Cultures ◽  
2D And 3D ◽  
3D Spheroids

Although 2D cell cultures are commonly used to predict therapy response, it has become clear that 3D cultures may better mimic the in vivo situation and offer the possibility of tailoring translational clinical approaches. Here, we compared the response of 2D and 3D colorectal cancer (CRC) cell lines to irradiation and chemotherapy. Classic 2D cultures and 3D spheroids of CRC cell lines (CaCo2, Colo205, HCT116, SW480) were thoroughly established, then irradiated with doses of 1, 4, or 10 Gy, using a clinical-grade linear accelerator. The response was assessed by immunohistochemistry, flow cytometry, and TUNEL assays. Upon irradiation, CRC 3D spheroids were morphologically altered. After irradiation with 10 Gy, annexin V/PI staining revealed a 1.8- to 4-fold increase in the apoptosis rate in the 2D cell cultures (95% CI 3.24±0.96), and a 1.5- to 2.4-fold increase in the 3D spheroids (95% CI 1.56±0.41). Irradiation with 1 Gy caused 3- and 4-fold increases in TUNEL positive cells in the CaCo2 and HCT116 (p = 0.01) 2D cultures, respectively, compared with a 2-fold increase in the 3D spheroids. Furthermore, the 2D and 3D cultures responded differently to chemotherapy; the 3D cultures were more resistant to 5-FU and cisplatin, but not to doxorubicin and mitomycin C, than the 2D cultures. Taken together, CRC cells cultured as 3D spheroids displayed markedly higher resistance to irradiation therapy and selected chemotherapeutic drugs than 2D cultures. This in vitro difference must be considered in future approaches for determining the ideal in vitro systems that mimic human disease.

scholarly journals Transient expression of interleukin 2 receptors. Consequences for T cell growth.

1983 ◽  
Vol 158 (6) ◽  
pp. 1895-1911 ◽  
Author(s):  
D A Cantrell ◽  
K A Smith
Keyword(s):  
Cell Proliferation ◽  
T Cells ◽  
T Cell ◽  
Cell Growth ◽  
Interleukin 2 ◽  
Flow Cytometric Analysis ◽  
Receptor Expression ◽  
T Cell Proliferation ◽  
Immune Stimulation ◽  
T Cell Growth

T lymphocyte mitosis results from the interaction of interleukin 2 (IL-2) with specific receptors that appear only after appropriate immune stimulation. To assess the potential role of IL-2 receptor levels in determining the rate and magnitude of T cell proliferation, the expression of IL-2 receptors by lectin-stimulated human peripheral blood T cells was examined and correlated with T cell growth. Using biosynthetically radiolabeled IL-2 and anti-Tac, a monoclonal antibody that blocks IL-2 receptor binding, IL-2 receptors were found to accumulate slowly and asynchronously among lectin-stimulated T cells and to precede the onset of DNA synthesis. Moreover, a critical threshold of IL-2 receptor density appeared to be required before the commitment to cell cycle progression, as analyzed quantitatively by tritiated thymidine incorporation and flow cytometric analysis of cellular DNA content. Once maximal IL-2 receptor expression occurred, continued proliferation was IL-2 concentration dependent as assessed using homogenous immunoaffinity-purified IL-2. Upon removal of the activating lectin, IL-2 receptor levels progressively declined, and, in parallel, the rate of proliferation diminished. The decay of IL-2 receptors could not be attributed to IL-2-mediated down-regulation. Instead, renewed IL-2 receptor expression was dependent upon the reintroduction of the initial activating signal. Repetitive exposure to lectin resulted in a more rapid reexpression of maximal IL-2 receptor levels, which was then followed by an accelerated resumption of proliferation. Thus, the extent of T cell proliferation after immune stimulation depends upon the interplay of the IL-2 concentration available and the density of IL-2 receptors expressed, both of which are ultimately determined by antigen/lectin stimulation. The awareness of the transience and the antigen/lectin dependence of IL-2 receptor expression, together with the capacity to monitor T cell cultures for IL-2 receptor levels, should facilitate the initiation and maintenance of cloned, antigen-specific T cells in long-term culture. In addition, these findings suggest that, in vivo, the rapidity of acquisition of maximum IL-2 receptor levels by activated T cells and the duration of IL-2 receptor expression may well direct the magnitude of T cell clonal expansion and resultant immune responses.

scholarly journals Comparison of senescence-related changes between three- and two-dimensional cultured adipose-derived mesenchymal stem cells

2020 ◽  
Author(s):  
Qiliang Yin ◽  
Na Xu ◽  
Dong sheng Xu ◽  
Ming xin Dong ◽  
Xiu min Shi ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Energy Metabolism ◽  
Cell Growth ◽  
Telomere Length ◽  
Cell Morphology ◽  
Telomerase Activity ◽  
Two Dimensional ◽  
3D Cultures ◽  
2D And 3D

Abstract Background: Adipose-derived mesenchymal stem cells ( ADMSCs ) have attracted widespread interest as cell-based tissue repair systems. To obtain adequate quantities of ADMSCs for therapeutic applications, extensive in vitro expansion is required. However, under current two-dimensional (2D) approaches , ADMSCs rapidly undergo replicative senescence , and cell growth is impeded and stem cell properties are eliminated by mechanisms that are poorly understood . These issues limit the extensive applications of ADMSCs . In this study, we investigated senescence-related changes in mesenchymal stem cells (MSCs) isolated from human adipose tissue in 2D and three-dimensional (3D) cultures. Methods: We studied cell growth over a given period (21 days) to determine if modes of culture were associated with ADMSCs senescence . ADMSCs were isolated from healthy females by liposuction surgery and then were grew in 2D and 3D cultures. The cell morphology was observed during cell culture. Every other time of culture, senescence-associated β-galactosidase (SA-β-gal) expression, cell viability, proliferation, and differentiation potential of ADMSCs from 2D and 3D cultures were detected. Also, senescence and stemness related genes expression, telomere length, telomerase activity, and energy metabolism of ADMSCs for different culture time were evaluated. Results: With long-term propagation, we observed significant changes in cell morphology, proliferation, differentiation abilities and energy metabolism, which were associated with increases in SA-β-gal activity, and decreases in telomere length and telomerase activity . Notably, when cultured in 3D, these changes were improved. Conclusions: Our results indicate that 3D culture is able to ameliorate senescence-related changes in ADMSCs.

Is Quercetin Beneficial for Colon Cancer? A Cell Culture Study, Using the Apoptosis Pathways

2021 ◽  
Vol 21 ◽  
Author(s):  
Meltem Özgöçmen ◽  
Dilek Bayram ◽  
İlkay Armağan ◽  
Gülçin Yavuz Türel ◽  
Murat Sevimli ◽  
...  
Keyword(s):  
Colon Cancer ◽  
Cell Proliferation ◽  
Effective Dose ◽  
Cell Lines ◽  
Caspase 3 ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Time Intervals ◽  
Cell Culture Study ◽  
3D Cultures ◽  
2D And 3D

Background: Quercetin (QCT) is a dietary flavonoid with many beneficial effects (e.g., antioxidant, antiaging, antidiabetic, antifungal effects, regulation of gastrointestinal motor activity in humans); furthermore, it induces apoptosis, cell cycle arrest, and differentiation. Objective: The apoptotic effects of OCT were investigated on SW480 human colon cancer cell lines in monolayer and spheroid cultures. Methods: Quercetin (40–200 μM) was applied, and inhibitory concentration (IC50) doses were determined for three-time intervals (24, 48, and 72 h). The effective dose was determined and applied for analyses, including staining with BrdU to investigate cell proliferation, terminal deoxynucleotidyl transferase dUTP nick, and labeling (TUNEL) to investigate apoptosis, and apoptosis-inducing factor (AIF) and Caspase-3 to investigate caspase-dependent or independent apoptotic pathways. Results: The effective dose of QCT was determined to be 200 μM and was found to induce apoptosis and inhibit cell proliferation at 24, 48, and 72 h, both in 2D and 3D cultures. Significant increases were observed in both caspase-3 and AIF staining, but cells showed greater caspase-3 staining compared with AIF staining at all time intervals (p<0.05). Conclusion: The QCT treatment groups showed more cell death and less cell growth than the untreated control groups in both 2D and 3D cultures of SW480 cell lines. The results suggest that quercetin induces apoptosis, inhibits cell proliferation, and has a protective role against colon cancer. However, further studies are needed to clarify its mechanism of action.

scholarly journals Transcriptome Analysis Reveals an Inhibitory Effect of Dihydrotestosterone-Treated 2D- and 3D-Cultured Dermal Papilla Cells on Hair Follicle Growth

2021 ◽  
Vol 9 ◽  
Author(s):  
Yufan Zhang ◽  
Junfei Huang ◽  
Danlan Fu ◽  
Zhen Liu ◽  
Hailin Wang ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
3D Model ◽  
Caspase 3 ◽  
Dermal Papilla ◽  
Inhibitory Effect ◽  
Follicle Growth ◽  
Hub Genes ◽  
Dermal Papilla Cells ◽  
Hair Follicle Growth ◽  
2D And 3D

Dermal papillae are a target of androgen action in patients with androgenic alopecia, where androgen acts on the epidermis of hair follicles in a paracrine manner. To mimic the complexity of the dermal papilla microenvironment, a better culture model of human dermal papilla cells (DPCs) is needed. Therefore, we evaluated the inhibitory effect of dihydrotestosterone (DHT)-treated two-dimensional (2D)- and 3D-cultured DPCs on hair follicle growth. 2D- and 3D-cultured DPC proliferation was inhibited after co-culturing with outer root sheath (ORS) cells under DHT treatment. Moreover, gene expression levels of β-catenin and neural cell adhesion molecules were significantly decreased and those of cleaved caspase-3 significantly increased in 2D- and 3D-cultured DPCs with increasing DHT concentrations. ORS cell proliferation also significantly increased after co-culturing in the control-3D model compared with the control-2D model. Ki67 downregulation and cleaved caspase-3 upregulation in DHT-treated 2D and 3D groups significantly inhibited ORS cell proliferation. Sequencing showed an increase in the expression of genes related to extracellular matrix synthesis in the 3D model group. Additionally, the top 10 hub genes were identified, and the expression of nine chemokine-related genes in DHT-treated DPCs was found to be significantly increased. We also identified the interactions between transcription factor (TF) genes and microRNAs (miRNAs) with hub genes and the TF–miRNA coregulatory network. Overall, the findings indicate that 3D-cultured DPCs are more representative of in vivo conditions than 2D-cultured DPCs and contribute to our understanding of the molecular mechanisms underlying androgen-induced alopecia.

scholarly journals Comparison of senescence-related changes between three- and two-dimensional cultured adipose-derived mesenchymal stem cells

2020 ◽  
Author(s):  
Qiliang Yin ◽  
Na Xu ◽  
Dong sheng Xu ◽  
Ming xin Dong ◽  
Xiu min Shi ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Energy Metabolism ◽  
Cell Growth ◽  
Telomere Length ◽  
Cell Morphology ◽  
Two Dimensional ◽  
Differentiation Potential ◽  
3D Cultures ◽  
2D And 3D

Abstract Background: Adipose-derived mesenchymal stem cells ( ADMSCs ) have attracted widespread interest as cell-based tissue repair systems. To obtain adequate quantities of ADMSCs for therapeutic applications, extensive in vitro expansion is required. However, under current two-dimensional (2D) approaches , ADMSCs rapidly undergo replicative senescence , and cell growth is impeded and stem cell properties are eliminated by mechanisms that are poorly understood . These issues limit the extensive applications of ADMSCs . In this study, we investigated senescence-related changes in mesenchymal stem cells (MSCs) isolated from human adipose tissue in 2D and three-dimensional (3D) cultures. Methods: We studied cell growth over a given period (21 days) to determine if modes of culture were associated with ADMSCs senescence. ADMSCs were isolated from healthy females by liposuction surgery and then were grew in 2D and 3D cultures. The cell morphology was observed during cell culture. Every other time of culture, senescence-associated β-galactosidase (SA-β-gal) expression, cell viability, and differentiation potential of ADMSCs from 2D and 3D cultures were detected. Also, senescence and stemness related genes expression, telomere length, telomerase activity, and energy metabolism of ADMSCs for different culture time were evaluated. Results: With long-term propagation, we observed significant changes in cell morphology, proliferation, differentiation abilities and energy metabolism, which were associated with increases in SA-β-gal activity, and decreases in telomere length and activity . Notably, when cultured in 3D, these changes were improved. Conclusions: Our results indicate that 3D culture is able to ameliorate senescence-related changes in ADMSCs.

Oncogenic LncRNA CASC9 in Cancer Progression

2020 ◽  
Vol 26 ◽  
Author(s):  
Yuying Qi ◽  
Chaoying Song ◽  
Jiali Zhang ◽  
Chong Guo ◽  
Chengfu Yuan
Keyword(s):  
Cell Proliferation ◽  
Drug Resistance ◽  
Cell Growth ◽  
Cancer Cells ◽  
Therapeutic Potential ◽  
Squamous Metaplasia ◽  
Neoplastic Transformation ◽  
Over Expression ◽  
Human Cancers ◽  
Current Review

Background: Long non-coding RNA (LncRNAs), with the length over 200 nucleotides, originate from intergenic, antisense, or promoter-proximal regions, is a large family of RNAs that lack coding capacity. Emerging evidences illustrated that LncRNAs played significant roles in a variety of cellular functions and biological processes in profuse human diseases, especially in cancers. Cancer susceptibility candidate 9 (CASC9), as a member of the LncRNAs group, was firstly found its oncogenic function in esophageal cancer. In following recent studies, a growing amount of human malignancies are verified to be correlated with CASC9, most of which are derived from the squamous epithelium tissue. This present review attempts to highlight the latest insights into the expression, functional roles, and molecular mechanisms of CASC9 in different human malignancies. Methods: In this review, the latest findings related to the pathophysiological processes of CASC9 in human cancers were summarized and analyzed, the associated studies were collected in systematically retrieval of PubMed used lncRNA and CASA9 as keywords. Results: CASC9 expression is identified to be aberrantly elevated in a variety of malignancies. The over-expression of CASC9 has been suggested to accelerate cell proliferation, migration, cell growth and drug resistance of cancer cells, while depress cell apoptosis, revealing its role as an oncogene. Moreover, the current review demonstrated CASC9 closely relates to neoplastic transformation of squamous epithelial cells and squamous metaplasia in non-squamous epithelial tissues. Finally, we discuss the limitations and tremendous diagnostic/therapeutic potential of CASC9 in various human cancers. Results: CASC9 expression is identified to be aberrantly elevated in a variety of malignancies. The over-expression of CASC9 has been suggested to accelerate cell proliferation, migration, cell growth and drug resistance of cancer cells, while depress cell apoptosis, revealing its role as an oncogene. Moreover, the current review demonstrated CASC9 closely relates to neoplastic transformation of squamous epithelial cells and squamous metaplasia in non-squamous epithelial tissues. Finally, we discuss the limitations and tremendous diagnostic/therapeutic potential of CASC9 in various human cancers. Conclusion: Long non-coding RNACASC9 likely served as useful disease biomarkers or therapy targets that could effectively apply in treatment of different kinds of cancers.

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