Profile of mouse Lewis lung carcinoma LLC1 cell line cultivated in 2-dimensional or 3-dimensional cell culture enriched with laminin rich extracellular matrix proteins
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ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE75864 | GEO | 2015/12/10
SECONDARY ACCESSION(S): PRJNA305546
REPOSITORIES: GEO
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Project description:This SuperSeries is composed of the SubSeries listed below. Refer to individual Series
2015-12-10 | E-GEOD-75864 | biostudies-arrayexpress
Project description:The extracellular matrix (ECM), as one of the key components of tumor microenvironment has a high impact on cancer development and highly influences tumor cell features. ECM contribute not only structural support to growing tumor cells, but also affect other cellular functions such as cell differentiation, migration, survival or proliferation. Moreover, gene and protein expression levels are regulated in cell-ECM interaction dependent manner. MicroRNAs (miRNAs) act as key modulators of gene expression in various basic cellular processes where they can decrease protein synthesis through translational repression or RNA degradation. Despite miRNAs have recently emerged as important players involved in regulation of cell-ECM interactions molecular mechanisms of this phenomenon remains to be elucidated. To address this question we explored miRNA expression patterns in mouse Lewis lung carcinoma LLC1 cells growing in laminin rich ECM microenvironment. Total RNA enriched in small noncoding RNAs was isolated 48h after growing in two different cell culture systems
2015-12-10 | E-GEOD-75862 | biostudies-arrayexpress
Project description:The extracellular matrix (ECM), as one of the key components of tumor microenvironment has a high impact on cancer development and highly influences tumor cell features. ECM contribute not only structural support to growing tumor cells, but also affect other cellular functions such as cell differentiation, migration, survival or proliferation. Moreover, gene and protein expression levels are regulated in cell-ECM interaction dependent manner. MicroRNAs (miRNAs) act as key modulators of gene expression in various basic cellular processes where they can decrease protein synthesis through translational repression or RNA degradation. Despite miRNAs have recently emerged as important players involved in regulation of cell-ECM interactions molecular mechanisms of this phenomenon remains to be elucidated. To address this question we explored miRNA expression patterns in mouse Lewis lung carcinoma LLC1 cells growing in laminin rich ECM microenvironment.
2015-12-10 | GSE75862 | GEO
Project description:Obvious advantages of 3D cell culture model are the cell morphology better reflecting tissue cell morphology, the formation of zones of i) active proliferation, ii) quiescent viable cell zone and iii) necrotic zone, as well as formation of nutrition, oxygen and drug gradients better reflecting cellular environment in tissue. Nevertheless the 3D cultures are a model still not resembling full complexity of tumor tissue environment in vivo. Few obvious limitations of 3D cell cultures as cancer research model are the lack of vasculature, host immune response and other cell-cell interactions that occur between cancer and stromal cells in tumors. Recognized advantages and limitations of the 3D cell culture models, however do not suggest directly the areas of cancer biology where 3D models could be applied with highest success. Hence detailed analysis at the molecular level of 2D/3D cell cultures and tumors in vivo are needed to unlock the power of 3D cell culture model. In order to elucidate which biological pathways of cancer cells in tumors are best resembled by the 3D cell culture model we have analyzed whole genome gene expression changes in mouse LLC1 cell line when cultured in 2D or laminin rich ECM 3D system. RNA was isolated 48h after growing in two different cell culture systems.
2015-12-10 | E-GEOD-75863 | biostudies-arrayexpress
Project description:Obvious advantages of 3D cell culture model are the cell morphology better reflecting tissue cell morphology, the formation of zones of i) active proliferation, ii) quiescent viable cell zone and iii) necrotic zone, as well as formation of nutrition, oxygen and drug gradients better reflecting cellular environment in tissue. Nevertheless the 3D cultures are a model still not resembling full complexity of tumor tissue environment in vivo. Few obvious limitations of 3D cell cultures as cancer research model are the lack of vasculature, host immune response and other cell-cell interactions that occur between cancer and stromal cells in tumors. Recognized advantages and limitations of the 3D cell culture models, however do not suggest directly the areas of cancer biology where 3D models could be applied with highest success. Hence detailed analysis at the molecular level of 2D/3D cell cultures and tumors in vivo are needed to unlock the power of 3D cell culture model. In order to elucidate which biological pathways of cancer cells in tumors are best resembled by the 3D cell culture model we have analyzed whole genome gene expression changes in mouse LLC1 cell line when cultured in 2D or laminin rich ECM 3D system.
2015-12-10 | GSE75863 | GEO
Project description:The extracellular matrix (ECM), one of the key components of tumor microenvironment, has a tremendous impact on cancer development and highly influences tumor cell features. ECM affects vital cellular functions such as cell differentiation, migration, survival and proliferation. Gene and protein expression levels are regulated in cell-ECM interaction dependent manner as well. The rate of unsuccessful clinical trials, based on cell culture research models lacking the ECM microenvironment, indicates the need for alternative models and determines the shift to three-dimensional (3D) laminin rich ECM models, better simulating tissue organization. Recognized advantages of 3D models suggest the development of new anticancer treatment strategies. This is among the most promising directions of 3D cell cultures application. However, detailed analysis at the molecular level of 2D/3D cell cultures and tumors in vivo is still needed to elucidate cellular pathways most promising for the development of targeted therapies. In order to elucidate which biological pathways are altered during microenvironmental shift we have analyzed whole genome mRNA and miRNA expression differences in LLC1 cells cultured in 2D or 3D culture conditions.In our study we used DNA microarrays for whole genome analysis of mRNA and miRNA expression differences in LLC1 cells cultivated in 2D or 3D culture conditions. Next, we indicated the most common enriched functional categories using KEGG pathway enrichment analysis. Finally, we validated the microarray data by quantitative PCR in LLC1 cells cultured under 2D or 3D conditions or LLC1 tumors implanted in experimental animals.Microarray gene expression analysis revealed that 1884 genes and 77 miRNAs were significantly altered in LLC1 cells after 48 h cell growth under 2D and ECM based 3D cell growth conditions. Pathway enrichment results indicated metabolic pathway, MAP kinase, cell adhesion and immune response as the most significantly altered functional categories in LLC1 cells due to the microenvironmental shift from 2D to 3D. Comparison of the expression levels of selected genes and miRNA between LLC1 cells grown in 3D cell culture and LLC1 tumors implanted in the mouse model indicated correspondence between both model systems.Global gene and miRNA expression analysis in LLC1 cells under ECM microenvironment indicated altered immune response, adhesion and MAP kinase pathways. All these processes are related to tumor development, progression and treatment response, suggesting the most promising directions for the development of targeted therapies using the 3D cell culture models.
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Project description:Profile of mouse Lewis lung carcinoma LLC1 cell line cultivated in 2-dimensional or 3-dimensional cell culture enriched with laminin rich extracellular matrix proteins
| PRJNA305546 | ENA
Project description:Conventional monolayer cell cultures used in cancer research accumulated basic knowledge of carcinogenesis which correlates with basic observations in tumor biology. Nevertheless, attempts to transfer accumulated cancer biology knowledge to clinical applications fails in most of the cases. Gathered data suggest that 2D cell culture models lack many features of tumor tissue in vivo. Although it has been demonstrated that cancer cells are generally more resistant to various types of treatments when cultured in extracellular matrix (ECM) based 3D cultures it is not clear if this phenomenon is attributed to the ECM dependent gain of function or the loss of non-physiological cellular state demonstrated while cultured at 2D conditions. To address the question of which universal molecular mechanisms if any are gained/lost in ECM-dependent manner we carried out genome wide gene expression analysis of human colorectal carcinoma DLD-1 and HT-29 cell lines cultured in 2D or ECM based 3D conditions.
2015-12-01 | GSE75551 | GEO
Project description:Although techniques for isolating and culturing adult cardiomyocytes were developed four decades ago, it still remains a challenge to isolate high yields of healthy viable cardiomyocytes, to maintain them in culture, and to use them successfully in experiments. This is due to the difficulty in deciding which adhesive substrate and buffer composition to use. Therefore this study aimed to (1) identify a robust experimental buffer to sustain survival of cultured adult rat cardiomyocytes (ARCMs) during control normoxic conditions, and (2) to identify an adhesive substrate that provides optimal cell adherence, not only during normoxia, but especially during simulated ischemia-reperfusion (SIR) experiments. Adhesion and viability of ARCMs were evaluated on laminin, laminin-entactin (LE), and extracellular matrix (ECM) at concentrations between 25-200 ug/ml, in three different normoxic experimental buffers and under SIR conditions. Differences in normoxic buffer composition had no effect on the adherence of ARCMs, but had a significant effect on mitochondrial function and thus cell viability. HEPES buffered PBS supplemented with 10 mM glucose was not sufficient to sustain cell viability unless 2 mM pyruvate was added, yet a cocktail of PBS and M199 provided an even greater viability. Finally, laminin, LE, and ECM retained similar numbers of ARCMs per concentration, but only provided efficient adhesion at concentrations ? 100 ug/ml.
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Project description:Full title: Comprehensive Characterization of Three-Dimensional Models for Prostate Cancer Growth and Invasion in Laminin-rich Extracellular Matrix Prostate Cancer (PrCa) cells undergo acinar morphogenesis and spheroid formation in three-dimensional (3D) culture, supported by laminin-rich extracellular matrix (lrECM, Matrigel). We developed miniaturized 3D model systems that facilitate investigation of morphogenesis and invasion of normal and PrCa cell lines in lrECM. Primary and non-transformed cell lines formed round structures with strong cell-cell contacts and epithelial polarization, lumen and a complete basal lamina (BL). In contrast, most PrCa cell lines formed either defective, “mass” spheroids with incomplete BL, or invasive “stellate” structures. The bioinformatic analyses of genome-wide mRNA expression data revealed massive alteration of key functional and signaling pathways in 3D cultures, with lipid and steroid metabolism, epigenetic reprogramming, and differentiation-related transcription factors induced across all cell lines by lrECM. In invasive cells, AKT, PI3Kinase, mTOR, and hedgehog signaling pathways were most highly activated, validated by small molecule inhibitors compounds specifically targeting key regulatory molecules. Compounds against AKT and PI3kinase pathways were significantly more effective in invasive cells, compared to mass or round/normal phenotype spheroids, and monolayer culture. A severe morphologic conversion was observed in PC-3 and PC-3M cells, transforming initially round, normal-appearing epithelial spheroids into rapidly invading cell masses. Markers for EMT (epithelial-mesenchymal transition) were highly expressed already in early stage, round spheroids prior to invasive conversion, and were not further increased in invasive cells. This indicates that PrCa cells can display extraordinary plasticity. EMT may be involved in providing a metastable genotype that allows morphological transformation, but is not be required for invasive processes themselves. Total RNA was obtained from non-transformed prostate epithelial cells and prostate cancer cells cultured in monolayer and three-dimensional laminin-rich extracellular matrix (growth factor-reduced Matrigel).
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