ABSTRACT: Conduct gene profiling experiment from a melanoma cell line which silenced galectin-3 gene expression to understand different phenotypic properties observed during tumor growth
Project description:Conduct gene profiling experiment from a melanoma cell line which silenced galectin-3 gene expression to understand different phenotypic properties observed during tumor growth.
Project description:Conduct gene profiling experiment from a melanoma cell line which silenced galectin-3 gene expression to understand different phenotypic properties observed during tumor growth. A melanoma cell line which silenced galectin-3 gene expression was obtained. This cell line was used in overexpression experiments, using a galectin-3 coding plasmid. Both galectin-3 negative and positive cells were obtained in syngeneic wild type and galectin-3 knockout mice. Altogether, we had evidence that while galectin-3 derived from the tumor cells impaired tumor growth, galectin-3 derived from stromal cells apparently favored tumor growth. Angiogenic response within tumors derived from galectin-3 expressing melanoma cells was delayed as compared to control conditions. Cells recruited to these distinct microenvironments seem to control tumor growth. Gene profiling of this tissue will allow for an integrative view of the process that was somehow orchestrated by galectin-3.
Project description:Malignant melanoma is a common and frequently lethal disease. Current therapeutic interventions have little effect on survival, emphasizing the need for a better understanding of the genetic, epigenetic, and phenotypic changes in melanoma formation and progression. We identified genes that were not previously known to be silenced by methylation in melanoma using a microarray-based screen following treatment of melanoma cell lines with the DNA methylation inhibitor 5-Aza-2'-deoxycytidine. Experiment Overall Design: RNA was isolated following 0 and 48 hours of 5AzadC treatment of melanocytes and six melanoma cell lines (MelJuSo, UACC 903, C8161, Neo6 C8161, WM1205, and WM35) and used for the reexpression microarray analysis.
Project description:To understand the transcriptional impact of FOXD3 in melanoma cells, we utilized a microarray approach. We collected RNA from three unrelated mutant B-RAF melanoma cell lines (WM115, WM793, and A375) that were engineered to inducibly express FOXD3 or the control gene, ?-galactosidase (LacZ), after 5 days of transgene induction. This time point was chosen based on maximal phenotypic changes previously observed. Comparison of gene signatures between the 3 cell lines produced approximately 2,600 common genes differentially regulated by FOXD3-expressing cells compared to the LacZ controls. Three unrelated mutant B-RAF melanoma cell lines (WM115, WM793, and A375) were induced to express FOXD3 and compared against the same cell lines expressing the control gene, ?-galactosidase (LacZ).
Project description:Although galectin-3 is known to modulate the cell proliferation, RNA processing, tumorigenesis, metastasis and apoptosis and also, which is highly expressed in human cancers, the function of galectin-3 is still controversy in gastric cancer. Here, we demonstrated the function of galectin-3 on gastric cancers by silencing with synthetic double-stranded small interfering RNA (siRNA). After silencing of galectin-3, cell numbers decreased and cell shapes changed in round. To determine the mechanism, microarray analysis was used to detect the changes in gene expression by galectin-3 silencing. we silenced galectin-3 in AGS as a gastric cancer cells with synthetic double-stranded small interfering RNA (siRNA), and performed the microarray analysis to detect the changes in gene expression by galectin-3 silencing.
Project description:Cellular stress contributes to the capacity of melanoma cells to undergo phenotype switching into highly migratory and drug tolerant dedifferentiated states. Such dedifferentiated melanoma cell states are marked by loss of melanocyte specific gene expression and increase of mesenchymal markers. Two crucial transcription factors, MITF and SOX10, important in melanoma development and progression have been implicated in this process. In this study we describe that loss of MITF is associated with a distinct transcriptional program, MITF promoter hypermethylation and poor patient survival in metastatic melanoma. From a comprehensive collection of melanoma cell lines, we observed that MITF methylated cultures were subdivided in two distinct subtypes. Examining mRNA levels of neural crest associated genes we found that one subtype had lost the expression of several lineage genes including SOX10. Intriguingly, SOX10 loss was associated with SOX10 gene promoter hypermethylation and distinct phenotypic and metastatic properties. Depletion of SOX10 in MITF methylated melanoma cells using CRISPR/Cas9 confirmed these findings. In conclusion, this study describes the significance of melanoma state and the underlying functional properties explaining the aggressiveness of such states.
Project description:The aim of the experiment was to study the effect on RNA processing upon Myg1 knockdown in B16 mouse melanoma cell line. Using siRNAs, Myg1 was silenced and the total RNA was isolated from defined compartments such as Nucleus and Mitochondria. From these samples, RNA sequencing analysis was performed to understand the changes in RNA number and RNA size.
Project description:To understand the transcriptional impact of FOXD3 in melanoma cells, we utilized a microarray approach. We collected RNA from three unrelated mutant B-RAF melanoma cell lines (WM115, WM793, and A375) that were engineered to inducibly express FOXD3 or the control gene, β-galactosidase (LacZ), after 5 days of transgene induction. This time point was chosen based on maximal phenotypic changes previously observed. Comparison of gene signatures between the 3 cell lines produced approximately 2,600 common genes differentially regulated by FOXD3-expressing cells compared to the LacZ controls.
Project description:This is the data to the melanoma-medium induction experiment. Tumor-associated and peripheral B cells from four patients were exposed to melanoma-conditioned medium (from one of the patients). This induced distinct phenotypic and functional changes in the B cells.
Project description:Prostate cancer (PCa) causes significant mortality among men worldwide. Galectin-4 plays an important roles in regulatiog cancer metastasis and high level of galectin-4 correlated with poor survival in subpopulation of PCa patients. To further understand the galectin-4-mediated signature and develop effective treatment, we froced expressed galectin-4 in 22Rv1 and LNCaP cells, and downregulated galectin-4 in 22Rv1-M4 cells. Next, cDNA microarray were applyed to study galectin-4-mediated gene expression and cancer progression.
