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Problems solved by technology
It is known that the expression of p53 is very important for this kind of cis-platinum to exert cytotoxicity on cancer cells (Apoptosis.2007Sep; 12(9):1733-42), according to different cell types, targeting cis-platinum The expression dynamics of ammonia-treated p53 are heterogeneous, and thus have the limitation of not highlighting the anticancer effect or consistent effect of cis-platinum
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Embodiment 1
[0102] Cisplatin and HPV E6 / E7 siRNA affect the effect of TP3 in cervical cancer cells
Embodiment 1-1
[0103] Effect of HPV E6 / E7 siRNA combined with cisplatin ammonia treatment in cervical cancer cells
[0104] Cervical cancer cells, HeLa cervical cancer cell line (HeLa; ATCC CCL-2) infected by HPV18 virus, and SiHa (SiHa; ATCC HTB-35) cervical cancer cell line infected by HPV16 virus by 5×10 4 or 1×10 5 The number of cells was put into a 6-well experimental plate, and placed in RPMI1640 or DMEM medium for 24 hours at 37°C and 5% carbon dioxide. The transfection of siRNA into cells was performed with the transfection reagent DharmaFect (Dharmacon, Lafayette, CO, USA) according to the instructions of the manufacturer.
[0105] In the case of Hela cervical cancer cell line infected by HPV18 virus, siRNAs of 426 and 450 sequences undergo shape-to-matter conversion individually and jointly, and in the case of SiHa cervical cancer cell line infected by HPV16 virus, siRNAs of 366, 44, and 497 sequences Transformation of shape and quality separately and jointly.
[0106] When usi...
Embodiment 1-2
[0110] Cell killing effect of cisplatin ammonia and HPV E6 / E7 siRNA
[0111] In order to confirm the effect of cisplatinum (CDDP) and HPV E6 / E7 siRNA on the viability of cervical cancer cells, the following experiments were performed.
[0112] Cervical cancer cells Hela and CaSki cells were transformed with HPV E6 / E7 siRNA, treated with 10 μM cisplatin, and cultured for 24 hours. After that, WST analysis was performed to confirm the viability of the cells.
[0113] The cell number was measured using the water-soluble tetrazolium salt (WST) method (EZ-Cytox kit; Daeil Institute of Analytical Research, Seoul, Korea). Put EZ-Cytox solution (50 μl) into each well of a 12-well experimental plate, and after incubating for 2 to 3 hours, use a GENios microplate reader (Tecan Trading AG, Wennidorf, Switzerland) to measure (485nm) viable cells.
[0114] The result is as Figure 1B As shown, compared with the non-cisplatin-treated population, the cell viability of the treated populat...
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Abstract
The present invention relates to a method for maintaining the increased intracellular p53 level, induced by a platinum-based anticancer drug, and an application thereof and, more specifically, to a method for maintaining the increased intracellular p53 level in cells by administering a platinum-based anticancer drug and siRNA to ubiquitin ligase for p53 to a subject in need thereof in combinationand sequentially, and to a composition for promoting cancer cell apoptosis using the same. According to the method of the present invention, the increased intracellular p53 expression level can be maintained for a long period of time in spite of the treatment with a low-concentration platinum-based anticancer drug, thereby effectively inducing the apoptosis of cancer cells and minimizing the drugside effect caused by the administration of the platinum-based anticancer drug, and thus the present invention can be favorably used in the prevention of cancer or the development of cancer medicines.
Description
technical field [0001] The application of the present invention claims the priority of US Patent Application No. 62 / 148,403, which was patented on April 16, 2015, and all contents of the specification are used as references of this patent application. [0002] The present invention relates to a method for maintaining the increased p53 gene level induced by platinum-based anticancer agents in cells and its application. Specifically, the composition involved is a combination of platinum-based anticancer agents and p53 ubiquitin ligase (ubiquitinligase) ) siRNA combined or sequentially administered to individuals in need, to maintain the method of increasing the level of p53 in cells and to use it to promote the killing of cancer cells. Background technique [0003] The tumor suppressor protein p53 can regulate the expression of multiple arrays of genes responsible for DNA repair, cell cycle arrest and growth and apoptosis, and plays a central role in maintaining genome integri...
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