Description
SCFSkp2-IN-2 (Compound AAA-237) is a Skp2 inhibitor with a K D of 28.77 µM. AAA-237 induces apoptosis of NSCLC cells and shows antitumor activitiesIn VitroSCFSkp2-IN-2 (Compound AAA-237) (0.3-3 µM; 24-72 h) can directly bind to Skp2, causing the degradation of Skp2 by proteasome. AAA-237 has no significant effect on the mRNA expression of Skp2 but inhibits the protein expression of Skp2 in NSCLC cells in a time-dependent manner. SCFSkp2-IN-2 (0.3-3 µM; 24-72 h) inhibits the proliferation of A549 and H1299 cells in a dose- and time-dependent manner. SCFSkp2-IN-2 (0.3-3 µM; 24-72 h) arrests the cell cycle at the G0/G1 checkpoint by regulating the Skp2-Cip/Kip and PI3K/Akt-FOXO1 signaling pathways. SCFSkp2-IN-2 (0.3-3 µM; 24-72 h) induces apoptosis of A549 and H1299 cells in a dose- and time-dependent manner. SCFSkp2-IN-2 (0.3 µM; 1-7 days) induces cellular senescence of the NSCLC cells. MCE has not independently confirmed the accuracy of these methods. They are for reference only. Western Blot AnalysisCell Line: A549 and H1299 cells Concentration: 0.3, 1 and 3 µM Incubation Time: 24, 48 and 72 h Result: Increased the expression levels of p21Cip1 and p27Kip1. Decreased the level of Skp2. Reduced the expression levels of CDK2, p-CDK2, cyclin E1, CDK4, p-CDK4 and cyclin D. Increased the levels of cleaved PARP, cleaved caspase 3, cleaved caspase 9 and Bax. Decreased the expression of PARP and Bcl-2. Cell Proliferation AssayCell Line: A549 and H1299 cells Concentration: 0.3, 1 and 3 µM Incubation Time: 24, 48 and 72 h Result: Inhibited the proliferation of A549 and H1299 cells in a dose- and time-dependent manner. The IC 50 for A549 was 3 µM at 24 h, 2.5 µM at 48 h and 0.7 µM at 72 h. The IC 50 for H1299 was 3.9 µM at 24 h, 1.8 µM at 48 h and 1.1 µM at 72 h. Cell Cycle AnalysisCell Line: A549 and H1299 cells Concentration: 0.3, 1 and 3 µM Incubation Time: 24, 48 and 72 h Result: Arrested the cell cycle at G0/G1 in a dose- and time-dependent manner. Apoptosis AnalysisCell Line: A549 and H1299 cells Concentration: 0.3, 1 and 3 µM Incubation Time: 24, 48 and 72 h Result: The ratio of JC-1 monomer/JC-1 polymer was increased in A549 and H1299 cells as the concentration was increased.In VivoSCFSkp2-IN-2 (Compound AAA-237) (15 or 45 mg/kg; i.p.; daily for 14 days) shows antitumor activity in lung cancer A549 xenograft mice model. MCE has not independently confirmed the accuracy of these methods. They are for reference only. Animal Model: A549 xenograft model Dosage: 15 or 45 mg/kg Administration: Intraperitoneal administration, daily for 14 days Result: Significantly reduced tumor volume and weight. The tumor growth inhibition of the low dose and high dose was 55% and 64%, respectively. No obvious weight loss or abnormal behavior was observed. Reduced the expression of Ki67 in tumor tissue. Reduced the expression of Skp2 and Bcl-2 and increased the expression of p27, the cleaving of caspase 3, caspase 9, PARP and Bax.IC50& Target:KD: 28.77 µM (Skp2)