MicroRNA expression profiles in head and neck cancer cell lines
Section snippets
Materials and methods
Cell lines and tissue culture. Investigations were carried out on nine mammalian cell lines. Details of the cell lines and their corresponding tissue culture techniques are presented in Table 1.
Detection of specific miRNA genes by RNA analysis. RNA was isolated from cell lines using Trizol (Invitrogen™, USA) as recommended by the manufacturer. Twenty micrograms of total RNA was electrophoresed in a 15% TBE–Urea gel using 1× RNA loading buffer (Ambion, USA). Following electrophoresis the RNA was
Relative expression of miRNA genes between head and neck cancer cell lines
The profiling data represented nine cell lines from four different anatomical head and neck sites. A reference set (Cy5-labelled) composed of oligonucleotides complementary to each miRNA probe on the array slide was used for the comparative analyses. Each array was co-hybridized with the reference set and sample (Cy3-labelled). In principle, the reference set acts as an internal hybridization control for every probe on the array. Expression of each miRNA was then calculated as a Cy3/Cy5 ratio
Acknowledgments
The study was funded by grants from The Sydney Cancer Foundation and The University of Sydney Cancer Fund.
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