Method of treating cancer using a neuropeptide Y5R (NPY5R) antagonist
一种癌症、低级的技术，应用在抗炎剂、抗肿瘤药、医药配方等方向，能够解决有待阐明等问题
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[0240] Example 1: siRNA silencing of the NPY Y5 receptor
[0241] Table 1. Loss of NPY5R expression
[0242]
example 2
[0243] Example 2: Cell Cycle Analysis
[0244] Breast cancer cell line MDA-MB-468 and lung cancer cell line A549 were purchased from ATCC. Cells were grown in medium recommended by ATCC, and the day before siRNA transfection a 6-well plate (2x10 per well 5 ) were plated in medium with 5.6 mM glucose.
[0245] The NPY5R siRNA pool was mixed from 4 individual duplexes purchased from Dharmacon. siRNA transfection was performed using Lipofectamine 2000 (Invitrogen) following the Invitrogen siRNA Transfection Scientific Pilot Program. A final 4OnM of siRNA was transfected into cultured cells.
[0246] Cells were harvested 3 days after siRNA transfection, washed once with PBS-0.1% BSA, and fixed with 75% ethanol-PBS-0.1% BSA at 40°C for at least 1 hour. - After washing twice with PBS-0.1% BSA, cells were stained overnight with 50 ug / ml propidium iodide and 10 ug / ml RNaseA in PBS. The next day cell cycle analysis was performed by flow cytometry ( figure 2 and image 3 ).
example 3
[0247] Example 3: Expression of NPY5R in various cancer cell lines
[0248] Expression of the Y5 receptor in cancer cell lines derived from several major cancer cell types (breast, lung, colon, prostate, and brain) was examined by Western blot ( Figure 4 ). High levels of NPY5R protein were detected in all cancer cell lines examined, consistent with its potential role in a wide range of cancers.
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