Introduction
Early childhood cancers are devastating diseases. There are several types of cancers that afflict children, but neuroblastoma ranks as the most prevalent and the most deadly. Approximately 1 out of every 8,000 live births will develop a form of neuroblastoma, consisting of roughly 9% of childhood cancers (Memorial Sloan Kettering Cancer Center, n.d.). The majority of children diagnosed with this form of cancer are around the ages of 1-2 where neuroblastoma is responsible for about 15% of all childhood cancer deaths (Thiele, 1998) Currently, there is no effective treatment option available for neuroblastoma patients. Therefore, there is an urgent need to perform neuroblastoma research to increase our understanding of the disease and to develop treatments for early childhood cancers.
There is ongoing research to find a cure for neuroblastoma. Recent reports have suggested that forced differentiation of tumor cells may serve as a promising strategy of neuroblastoma therapy. Chemical compounds such as retinoids, which are structurally similar to vitamin A, are frequently used as cell differentiation inducers. Retinoids such as all-trans-retinoic-acid (ATRA) induce differentiation of target cells by acting on cell signaling pathways and activating nuclear receptors proteins RAR and RXR (Wikipedia, 2014). ATRA-activated nuclear receptors directly alter gene expression resulting in cell differentiation by inducing vitamin A which is essential in the differentiation of a neuron. The potential of retinoids in the treatment of various types of cancer cells is intriguing and promising. Unfortunately, toxicity and acquired resistance to retinoids limits their clinical use because of its effect on differentiated cells.
Recently, a paper published by researchers at Johns Hopkins University shows that any cell has the ability to return to a stem cell if the correct properties are exerted. This study applies a similar strategy by applying retinoic acid to undifferentiated neuroblastoma cells to force the cell to become a differentiated neuron. In this experiment retinoic acid is used to force these undifferentiated cancer cells to produce the proteins responsible for allowing electron transfer. Likewise, the retinoic acid should induce morphological changes and changes in gene expression that will lead to functional changes in the cell. This research paved the way for other possibilities in the alteration of gene expression and is important to the research reported in this paper.
SKNSH cells have been shown to differentiate upon treatment with retinoic acid. When these cells are treated with retinoic acid they are able to express what was a repressed gene due to a previous mutation. When treated with retinoic acid these repressed genes are able to be turned on and the cell is able to continue its development into a differentiated neuron. GFAP and β-Tubulin are two of the proteins associated with neuronal differentiation. When treated with retinoic acid the expectation is that the neuroblastoma cells are able to become functional neurons and express outgrowths, or projections, that will allow the neuron to transfer electrical signals which is indicative of a neuron. Also, the hope is to see qualitative data that shows morphological changes in the treated cells.
The investigation will look at the hypothesis that neuroblastoma cells treated with retinoic acid will undergo cell differentiation through changes in gene expression and acquire distinct morphological characteristics. In this research project cells were treated with retinoic acid over a set period of time and consequential tests were performed to measure phenotypically changes. Specifically, we treated the SK-N-SH neuroblastoma cell-line with retinoic acid over a time-course and measured the consequential phenotypic changes using confocal microscopy. Several cell components were investigated including actin, GFAP, and β-tubulin to form detailed conclusions about morphological and gene expression changes that accompany neuroblastoma differentiation. Also, conductivity was examined in order to determine the functional changes of the cell after treatment. Because the expression of many genes are unregulated in neuroblastoma the differentiation of the cell may give rise to the neuronal phenotype associated with retinoic acid treatment. The collected results will in theory, consequently define the cell as a differentiated neuron rather than a neuroblastoma cancer thus considering the cancer cured.
Early childhood cancers are devastating diseases. There are several types of cancers that afflict children, but neuroblastoma ranks as the most prevalent and the most deadly. Approximately 1 out of every 8,000 live births will develop a form of neuroblastoma, consisting of roughly 9% of childhood cancers (Memorial Sloan Kettering Cancer Center, n.d.). The majority of children diagnosed with this form of cancer are around the ages of 1-2 where neuroblastoma is responsible for about 15% of all childhood cancer deaths (Thiele, 1998) Currently, there is no effective treatment option available for neuroblastoma patients. Therefore, there is an urgent need to perform neuroblastoma research to increase our understanding of the disease and to develop treatments for early childhood cancers.
There is ongoing research to find a cure for neuroblastoma. Recent reports have suggested that forced differentiation of tumor cells may serve as a promising strategy of neuroblastoma therapy. Chemical compounds such as retinoids, which are structurally similar to vitamin A, are frequently used as cell differentiation inducers. Retinoids such as all-trans-retinoic-acid (ATRA) induce differentiation of target cells by acting on cell signaling pathways and activating nuclear receptors proteins RAR and RXR (Wikipedia, 2014). ATRA-activated nuclear receptors directly alter gene expression resulting in cell differentiation by inducing vitamin A which is essential in the differentiation of a neuron. The potential of retinoids in the treatment of various types of cancer cells is intriguing and promising. Unfortunately, toxicity and acquired resistance to retinoids limits their clinical use because of its effect on differentiated cells.
Recently, a paper published by researchers at Johns Hopkins University shows that any cell has the ability to return to a stem cell if the correct properties are exerted. This study applies a similar strategy by applying retinoic acid to undifferentiated neuroblastoma cells to force the cell to become a differentiated neuron. In this experiment retinoic acid is used to force these undifferentiated cancer cells to produce the proteins responsible for allowing electron transfer. Likewise, the retinoic acid should induce morphological changes and changes in gene expression that will lead to functional changes in the cell. This research paved the way for other possibilities in the alteration of gene expression and is important to the research reported in this paper.
SKNSH cells have been shown to differentiate upon treatment with retinoic acid. When these cells are treated with retinoic acid they are able to express what was a repressed gene due to a previous mutation. When treated with retinoic acid these repressed genes are able to be turned on and the cell is able to continue its development into a differentiated neuron. GFAP and β-Tubulin are two of the proteins associated with neuronal differentiation. When treated with retinoic acid the expectation is that the neuroblastoma cells are able to become functional neurons and express outgrowths, or projections, that will allow the neuron to transfer electrical signals which is indicative of a neuron. Also, the hope is to see qualitative data that shows morphological changes in the treated cells.
The investigation will look at the hypothesis that neuroblastoma cells treated with retinoic acid will undergo cell differentiation through changes in gene expression and acquire distinct morphological characteristics. In this research project cells were treated with retinoic acid over a set period of time and consequential tests were performed to measure phenotypically changes. Specifically, we treated the SK-N-SH neuroblastoma cell-line with retinoic acid over a time-course and measured the consequential phenotypic changes using confocal microscopy. Several cell components were investigated including actin, GFAP, and β-tubulin to form detailed conclusions about morphological and gene expression changes that accompany neuroblastoma differentiation. Also, conductivity was examined in order to determine the functional changes of the cell after treatment. Because the expression of many genes are unregulated in neuroblastoma the differentiation of the cell may give rise to the neuronal phenotype associated with retinoic acid treatment. The collected results will in theory, consequently define the cell as a differentiated neuron rather than a neuroblastoma cancer thus considering the cancer cured.