| Popis: |
BACKGROUND: This study is devoted to the search for molecular features of human glioblastoma cells with high migration abilities, with a special focus on the nucleolin, as a marker. Nucleolus disposed protein, nucleolin, is highly expressed in malignant tumor cells. Its localization in the cytoplasm and on the cell membrane is considered to be a poor prognostic indicator. Particular attention was paid to a significance of the intact C-terminus of nucleolin for distribution within the cells of transplanted human glioblastoma cultures derived from patient tissues. MATERIAL AND METHODS: Several primary cultures derived from human gliomas GrIV with different migration properties were used in this study. The methods of studying are: qRT-PCR, cloning, sequencing, rapid amplification of cDNA Ends, Southern blot hybridization of products of 3’RACE and 2D Western Blot. RESULTS: Using fused nucleolin with labeled C-and N-terminals, it has been shown that in glioblastoma cells with a high degree of invasion nucleolin is located in the cytoplasm and close to cell membrane, and the distribution of nucleolin with modified either C- or N-terminal is different. CONCLUSION: This study shows importance of the intact C-terminus of nucleolin for its conventional nuclear localization. At the same time, we suggest that a change in the C-terminus of nucleolin, as well as movement of this protein into cytoplasm and into membrane could be characteristic of glioblastoma cells with a high degree of invasion. The reported study was funded by RFBR according to the research project №17-00-00157, № 17-00-00158 (17-00-00162 (K)) and 18-015-00279. |
