AICAR CAS:2627-69-2 AMPK activator High Purity
AICAR CAS:2627-69-2 AMPK activator High Purity
Our results provided new and informative clues for better understanding of the role of AMPK in β-cell apoptosis. The biguanide drug metformin Cipandrol (Testosteron C) 200 mg Balkan Pharmaceuticals buy online has anti-tumor activity 21 and patients taking this drug to treat their type 2 diabetes may have a reduced risk of certain cancers 27, including prostate cancer 28. Although the cancer killing effect of metformin has been attributed to the indirect activation of AMPK 29, anti-proliferative effects of metformin in LNCaP cells have also been proposed to occur in an AMPK-independent manner 30.
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The effect of each compound on each cell on each of the above parameter is presented in Fig. For example, bezafibrate increased growth in C20ORF7 approaching that in GLU medium. On the contrary, genistein, EGCG and grape seed extract had a negative effect on growth.
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After 24 h incubation, MTT was added to a final concentration of 0.5 mg/ml and cultures were incubated for 2 h. To assess the cellular apoptosis, we used Annexin V-FITC Apoptosis Detection Kit (Abcam) according to the manufacturer’s protocol. Then, 5 × 105 cells were collected and incubated with Annexin V-FITC for 5 min at room temperature in the dark. Afterward, the cells were detected by flow cytometry using Flow Cytometer laser 488 nm (Becton Dickinson, NJ) and analyzed with FlowJo™ Software 31.
As shown in Figure 1C, apoptosis and autophagy were not activated in dose- and time-dependent manner when the cells were treated with metformin. For further confirmation, all these treated cells were analyzed by electron microscopy and flow cytometry. These three cells displayed extensive apoptotic cells (Figure 2A) and an increased sub-G1 population (Figure 2B) after AICAR treatment. As seen in Figure 2B, we observed the same percentage of cells in G0/G1, G2and S phase in metformin treated cells compared to their controls. Taken together, our data demonstrate that metformin did not induce apoptosis, autophagy and cell cycle arrest. The cytotoxicity of AICAR was apparent in two prostate cell lines, PC3 and LNCaP, in two in vitro models of tumor growth, clonogenic assays using 2-dimensional cultures and multicellular tumor spheroids using 3-dimensional cultures.
- To investigate the intracellular signal transduction system in KGN cells, we performed a Western immunoblotting analysis as described 25.
- It was reported that AMPK suppressed the TNFα-induced cytokine production in other cells 32.
- FANCD2 activation was abolished by treatments with Compound C, an AMPK inhibitor, or after AMPKalpha1 knockdown, substantiating the involvement of AMPK in AICAR-induced FANCD2 activation.
Cell Culture
Then, the MTT solution was removed, and 200 μl of DMSO (Merck) was added to each well. The optical densities (ODs) of the stained solutions were measured with POLARstar Omega Plate Reader Spectrophotometer at 570 nm wavelength 29. Mesenchymal stromal cell (MSC) stemness capacity diminishes over prolonged in vitro culture, which negatively affects their application in regenerative medicine. To slow down the senescence of MSCs, here, we have evaluated the in vitro effects of 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR), an AMPK activator, and nicotinamide (NAM), an activator of sirtuin1 (SIRT1). In conclusion, we provide evidence for both AMPK and PGC-1α in regulating protein abundance of SIRT3 and MnSOD.