ABSTRACT
Objective: To screen the anti-atherosclerosis (AS) activity of the compounds by using THP-1-HIF-1α-HER-Luciferase high-throughput model, and to verify the anti-AS function of the effective compounds. Methods: THP-1-HIF-1α-HER-Luciferase cells were pretreated with different concentrations of compounds (1, 10, and 100 μg/mL) for 2 h, then cultured under hypoxia for 24 h. Luciferase activity of cells was detected and compounds with anti-AS activity were screened by luciferase activity evaluation. THP-1 and U937 cells were pretreated with effective compounds, and then induced for 24 h by oxidized low density lipoprotein (OX-LDL). The formation of foam cells was observed by oil red staining. The mRNA level of hypoxia-inducible factor 1α (HIF-1α) was detected by real-time quantitative PCR (qPCR). HIF-1α protein expression was detected by Western blotting. Anti-AS activity of effective compounds were evaluated. Results: Among the 200 compounds, 11 compounds could significantly inhibit the increase of luciferase activity in THP-1-HIF-1α-HER-Luciferase cells induced by hypoxia (all P<0.05), and compound numbered 14 (C14) had the most significant inhibitory effect. THP-1 and U937 cells formed foam cells induced for 24 h by OX-LDL. However, cells were pretreated with C14 for 2 h, which could significantly inhibit the formation of foam cells induced by OX-LDL. Cells were induced for 24 h by OX-LDL, which could significantly increase the expression of HIF-1α mRNA and protein (all P<0.05), while cells pretreated with C14 could significantly inhibit the increase of HIF-1α mRNA and protein expression in a gradient-dependent manner (all P<0.05). Conclusion: THP-1-HIF-1α-HER-Luciferase high-throughput model can be reliability used in screening of compounds with anti-AS activity. C14 has the good anti-AS activity characteristics.
ABSTRACT
<p><b>OBJECTIVE</b>To study the effect of LY294002 on the adriamycin- induced epithelial-mesenchymal transition in human breast carcinoma cells.</p><p><b>METHODS</b>Human breast carcinoma cells MCF-7 was cultured in vitro and then exposed to adriamycin with or without LY294002. The protein expression levels of Akt, phosphorylated-Akt (p-Akt), Snail, and E-cadherin was detected by Western blot analysis. The mRNA expressions of Snail and E-cadherin were determined by RT-PCR.</p><p><b>RESULTS</b>Adriamycin significantly increased the protein expression of Snail and depressed the protein expression of E-cadherin (P<0.05). The pre-treatment with LY294002 significantly reversed the changes of activities and levels of the above proteins (P<0.05).</p><p><b>CONCLUSION</b>LY294002 could reverse the adriamycin-induced epithelial-mesenchymal transition in human breast carcinoma cells by regulating the expressions of Snail and E-cadherin through suppressing PI3K/Akt signaling pathway.</p>