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The Antitmor Activity of Bis(6-hydroxy-4-oxo-4H-chromene-3-yl)-ethylenediimine at HeLa Cells by Upregulated p53 Protein

Author: CaiTiKuan
Tutor: WangQin
School: Lanzhou University
Course: Biophysical
Keywords: Hela Human papillomavirus Proto-oncogene p53 MDM-2 Western blot
CLC: R737.33
Type: Master's thesis
Year: 2007
Downloads: 45
Quote: 0
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Abstract


Background: The human papilloma virus (Human Papillomavirus, HPV) infection is a major cause of cervical cancer. HPV encoded E6 protein and p53 protein and intracellular E6-associated protein (E6 associated protein, E6-AP) combined with p53 degradation through the ubiquitin pathway, so that the normal cells become cancerous, the formation of malignant tumors. active role in p53 cells maintain genetic stability, which is mainly reflected in the cell cycle arrest, promoting apoptosis and DNA repair. MDM2 protein is intracellular adjusting the concentration and activity of the p53 protein, both of which constitute similar current loop negative feedback to each other for fine adjustment. MDM2-mediated p53 degradation and direct inhibition of p53 transcriptional activity are two forms of regulation of p53 function through the nuclear membrane into the cytoplasm. Flavonoids (flavonoids compounds) plant secondary metabolites, exists widely in natural plants, type of structure is complex and diverse, exhibit a variety of pharmacological activities, the system of prevention and treatment of cardiovascular and cerebrovascular diseases and respiratory diseases, have anti-inflammatory antibacterial, lowering blood sugar, anti-oxidation, anti-radiation, anti-cancer, anti-tumor and enhance immunity pharmacological effect. Flavonoids has become a hotspot of drug development. Methods: The newly synthesized flavonoids Schiff base of 2 - (6 - hydroxy-4 - oxo-4H-benzopyran -3 - yl) - ethylenediamine (bis (6-hydroxy-4-oxo-4H -chromene-3-yl)-ethylenediimine, B6) in the treatment of Hela cells with different concentrations at different times. By SRB, trypan blue staining, detection of the inhibitory effect of the compound on Hela cells; observed changes in cell morphology caused by the action of the compound under phase contrast microscope; morphology of apoptotic cells was observed by AO / EB double staining; agarose gel electrophoresis, the compound effect of cell DNA to form bands; TEM observation cell formation of apoptotic bodies; flow cytometry Hela cell apoptosis and cell cycle arrest; Western Blot detection of p53 of MDM2 protein expression changes . Results: SRB assay to 24h Hela cells IC 50 43.51μmol / L; trypan blue staining detected compounds 24h Hela cells significantly inhibited; difference between the microscope and the AO / EB ultraviolet excitation of the double staining were observed under a fluorescence microscope to the cell membrane separation, cell apoptosis corpuscles; can be observed under the ultraviolet excitation of agarose gel electrophoresis of DNA ladder strip is formed at 180 ~ 200bp between; see TEM cells chromatin condensation, separation and distribution along the nuclear membrane the cytoplasm also occur pyknosis, apoptosis precursors; flow cytometry cells arrest in G1 phase, apoptosis and apoptosis rate in the 0 μmol / L, 40μmol / L, when the concentration of 80μmol / L, respectively, 0.05%, 6.1%, 8.6%; increase the Western Blot p53 protein increased with increase in concentration of compound that increases the amount of the amount of MDM2 protein with the p53 protein. The above results indicate that the compound is capable of Hela cells induced to undergo apoptosis, and p53 expression promoting promote apoptosis, is a compound having anti-tumor activity.

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CLC: > Medicine, health > Oncology > Genitourinary tumors > Female genital tumors > Uterine tumors
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