Expression of Bcl-2 and other regulatory genes in primary cell culture derived from spinocellular tumour of head and neck
|Year of publication
|Relative lack of symptoms of head and neck tumours together with limited effect of treatment of advanced stages highlights the need of novel cancer biomarkers. Consistently with the identification of such molecules, understanding of molecular mechanisms of head and neck tumours is needed. Therefore, we prepared a primary cell culture from advanced spinocellular pharyngeal tumour and analysed the expression of apoptosis- and cell cycle-related genes. Following genes were analysed: Bax, Bcl-2, p53. The expression of these genes was analysed on qRT-PCR using Taqman probes and relative ddCt method, and standardized to beta-actin as a housekeeping gene. To eliminate the effect of fibroblast overgrowing, medium with D-valin was used. Non-tumour tissue (not treated with D-valin) was used as a relative control. Additionally to qRT-PCR, cell growth was analysed using impedance-based real-time cell growth monitorring system. In this pilot study, we determined significant differences in the expression of both Bax and Bcl-2 and no significant difference in the expression of p53 gene (p < 0.05, t-test). These preliminary data suggest this particular cell culture of head and neck utilize inhibition of apoptosis as a way to promote tumorigenesis. regulation is distinctly affected in this primary cell culture. As a result of modification of these processes, differential growth of tumorous and non-tumour primary cell culture was determined. However, further experiments focused on the characterisation of this cellular culture are needed.