Differential mRNA Expression of AURKA ,AURKB and AURKC in Neuroblastoma Cell Lines
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Heng Liang
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Introduction
Aurora kinase family plays an important role in mitosis. Since their discovery in the 1990s, Aurora kinases have been strongly linked to the progression of human cancers. Aurora A maps to human chromosome 20q13 and Aurora B to human chromosome 17q13.1, which are loci altered frequently in human cancers. Overexpression of Aurora A and B is seen in many cancers. Aurora C is the least well studied member of this family and is expressed in most somatic tissues at levels much lower than Aurora A or B.
Neuroblastoma is derived from the arrested differentiation of neural crest progenitor cells. Recent study showed that Aurora kinase family has close relationship with neuroblastoma (Unholy Matrimony: Aurora A and N-Myc as MalignantPartnersinNeuroblastomaCancer Cell, Volume 15, Issue 1, 67-78, 6 January 2009).
This project investigated the expression levels of the AURKA,AURKB and AURKC genes, between N type (represented by KCN-83n) ,S type (represented by SH-EP1) and I type (represented by BE(2)-C) neuroblastoma cell lines which exhibit differences in their invasiveness properties.
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Figures
Figure 1-RT-PCR analysis of AURKA transcript levels in N,I and S cells. RT-PCR products generated from RNA isolated from each of the cell lines, amplified by primers recognizing GAPDH and AURKA respectively.
Figure 2-RT-PCR analysis of AURKB transcript levels in N, I and S cells. RT-PCR products generated from RNA isolated from each of the cell lines, amplified by primers recognizing GAPDH and AURKB respectively.
Figure 3--RT-PCR analysis of AURKC transcript levels in N,I and S cells. RT-PCR products generated from RNA isolated from each of the cell lines, amplified by primers recognizing GAPDH and AURKC respectively.
Figure 4- The sequence of the AURKA, AURKB and AURKC amplified using specific primers with a segment of Homo sapiens mRNA sequence from NCBI.
The aim of this project was to investigate the mRNA expression levels of the aurora kinase genes in N,I and S neuroblastoma cells. The RT-PCR analysis suggested a higher expression for AURKA mRNA in I, while Aurora C is expressed the lowest in the I cell type, as compared to N and S cellS. There appeared to be no significant difference in AURKB mRNA expression in the three cell phenotypes.
Consistent with results from RT-PCR (Fig. 1) of this study, several other investigators reported increased expression of AURKA mRNA in highly invasive cancer cells. However, the reason of decreased AURKC expression in I cells is not clear.
The partial sequence of each of the three aurora kinase products from RT-PCR amplified using primers specific to each kinase gene, confirmed that these primers amplified the desired aurora kinase transcript, respectively (Fig. 4).
Based on the obtained data, it cannot be concluded that, the increased expression of AURKA and decreased expression of AURKC have direct relation of the invasiveness of neuroblastoma cell. There are other factors that may also affect the invasiveness of these cells. Due to the increasing relevance of abberations in the aurora kinase dependent pathway in human tumorigenesis, further studies are warranted to study the level and activity of other components of this pathway . Research in this direction could prove useful to develop molecular therapeutics targeting Aurora Kinases.
Full Paper
Acknowledgments
I would like to thank Bo Liu and Leleesha Samaraweera for their help throughout the course. I would also like to thank Dr. Ross for providing neuroblastoma cells . Finally ,I would like to acknowledge Dr. Berish Rubin, and Dr. Sylvia Anderson for their guidance.
