Neuroblastoma is a malignant, solid tumor that develops from embryonic neural crest cells found in several areas of the body (most commonly originating in and around adrenal glands). It usually occurs in children younger than five years and sometimes forms before a child is born. It is slightly more common in males. (5)
Figure 1-Transcript Variants of VEGFA, Positions of primer sets 1F/1R and 2F/2R, Expected product lengths of each transcript variant with primer set 2F/2R
Figure 2-Primers 1F and 1R amplifies invariant section (between exon 3 and exon 4) of all transcript variants of VEGFA in all four neuroblastoma cell lines
Figure 3-Detection of two alternatively spliced variants of VEGFA with primers 2F and 2R in all four neuroblastoma cell lines. The levels of TV 4 and TV 6 are equivalent in both S and N cell lines studied. The level of TV 6 is relatively higher in both I type cell lines when compared with the levels of TV 4
Figure 4-GAPDH was used as a loading control
Neuroblastoma is a malignant, solid tumor that develops from neural crest cells and usually occurs in children younger than 5 years. The neuroblastoma cell lines exhibit 3 phenotypes – N ( Neuroblastic), S (Substrate adherent) and I (Intermediate). I type cells are the most malignant. The mRNA expression of VEGFA was studied in these cell lines. VEGFA is a signaling protein, a member of Platelet Derived Growth Factor (PDGF), associated with angiogenesis and vasculogenesis. Angiogenesis plays an important role in tumor development. Differential expression of VEGFA mRNA in a few neuroblastoma cell lines was found. Two transcript variants were expressed in all four cell lines. Transcript variant four and six were detected. No significant differences in the expression of these two transcript variants were found in the N and S cell lines. However, the levels of transcript variant six appeared to be higher in both I type cell lines studied when compared to transcript variant 4.
I would like to thank Dr. Berish Rubin for his guidance and giving me an opportunity to accomplish this project. I would also like to thank Alex and Xie Xie for their consistent support and patience throughout the project. Further I would like to thank Dr. Ross for generously providing neuroblastoma cell lines. Additional thanks to Dr. Sylvia Anderson, Bo Liu and Class of 2011 for their help and encouragement throughout the project.
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