Supervisors info:
Διαμάντης Σίδερης, Αναπληρωτής καθηγητής, Τμήμα Βιολογίας, Εθνικό και Καποδιστριακό Πανεπιστήμιο Αθηνών
Summary:
RNA molecules in biology systems are well-known to play a significant role in a variety of cell functions, from proteinosynthesis to cell regulation. Therefore, the maintenance of a perfect balance between their synthesis and degradation is essential, with ribonucleases playing that part. A recently described family of ribonucleases, the RNase κ family, has been a subject of study for our laboratory for the past few years. The human RNase κ was till now represented by 2 isoforms of the RNase κ gene in mRNA level. Experiments conducted by Next Generation Sequencing (NGS) technology have shown 17 possible new variants of the RNase κ gene in cancer cell lines. The aim of this research project was the research of the expression of these new variants in cancer cell lines and the study of their expression patterns. We focused on 10 out of the 17 new variants, which were studied by pairs, variant 2 and 4, variant 5 and 14, variant 6 and 17, variant 12 and 13 and variant 15 and 16. The protocol which was followed included the culture of cancer cell lines derived from 18 different cancer tissues, RNA extraction, reverse transcription, polymerase chain reaction (PCR) and nested PCR. In order to detect the new variants, we conducted agarose gel electrophoresis. The results verified the existence of these new variants in these cancer cell lines, which were expressed in diverse manner. There were variants such as variant 4 and 17 which were expressed in almost all cell lines whereas they were variants like 15 which was absent in most of all cell lines and other like variant 13 which was expressed in a significant quantity in specific cancer cell lines. Therefore, taking into consideration that the expression of alternative variants of proteins in different cancer types and their levels of expression have been used in Molecular Diagnostics field as cancer biomarkers, these results can be further used for the correlation of specific RNase κ variants with specific cancer types.
