The New Method Of Measuring The Level Of VEGF-R1 In The Nucleus Of A Cancer Cell Is The Possibility To Evaluate The Degree Of Cell Malignization
Objective of the study. The work studied VEGF-R1 and its role in the possibility of the evaluation of the degree of cancer cell malignization. Materials and Methods. Two lines of human tumor cells A431 and BRO were used as the objects of study. The line of normal human fibroblasts (PHF) served as a control line. The quality of extracted nuclei was assessed based on the criteria of their intactness and purification from cytoplasm. The presence of nuclear markers PCNA and lamin A/C, based on the data offlow cytometry (FC), was indicative ofpreservation of nuclear proteins and the absence of the cytoplasmic β - tubulin confirmed that that the nuclei did not contain cytoplasmic impurities. After implementation of controls, the content of VE-GF-R1 was measured by flow cytometry (FC). Results. The high content of PCNA and lamin A/C in the extracted nuclei with almost complete absence of β - tubulin proved that the protocol might be used to obtain a pure suspension of single intact cell nuclei. The measurement of the nuclear VEGF-R1 content revealed that it was present only in tumor cell nuclei, and that in more malignant BRO cells the receptor content was 1.75 times higher than in A431. Conclusions. The developed method of extraction of cell nuclei for their subsequent flow cytometry analysis was found to be suitable for the quantitative evaluation ofprotein content in the native nucleus. The presence of VEGF-R1 in the nuclei of tumor cells rather than in normal cells supports the assumption that the nuclear receptor contributes to carcinogenesis. The revealed tendency to increased content of the receptor in the nucleus as the malignancy of cell line increases indicates that the nuclear receptor ‘s level may be used for prognostic purposes. In addition, these results allow to consider VEGF-R1 itself as a potential target in anticancer therapy.