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OBJECTIVE： To study the effects of aspirin on the growth and autoghagy of human gastric cancer cells SGC-7901 and BGC-823. METHODS： SGC-7901 and BGC-823 cells were selected as research objects， with phosphate buffer （PBS） as negative control treated for 48 h， MTT assay was used to detect the effects of 1， 2， 4， 6， 8， 10 mmol/L aspirin， 5 mmol/L aspirin alone or combined with 2.5 μmol/L chloroquine， 2.5 μmol/L 3-methyladenine （3-MA） on survival rate of gastric cancer cells. Flow cytometry was used to detect the effects of 2 and 5 mmol/L aspirin， 5 mmol/L aspirin alone or combined with 2.5 μmol/L chloroquine and 2.5 μmol/L 3-MA on the apoptosis rate and cell cycle distribution of gastric cancer cells. Hoechst33258 staining was used to observe the effects of 5 mmol/L aspirin on morphology of gastric cancer cell nucleus； Transwell chamber test was adopted to detect the effects of 5 mmol/L aspirin on the migration of gastric cancer cell. Laser confocal scanning microscopy was used to observe the effects of 5 mmol/L aspirin on autophagy formation in gastric cancer cells. Western blot method was used to detect the effects of 2 and 5 mmol/L aspirin on the protein expression of autophagy markers LC3-Ⅱin gastric cancer cells. RESULTS： Compared with negative control group， aspirin could inhibit the survival rates of SGC-7901 and BGC-823 cells in dose-dependent manner， but had no significant effects on apoptosis rate of SGC-7901 and BGC-823 cells； SGC-7901 and BGC-823 cells were blocked in G1 phase. Compared with aspirin alone group， the survival rates of SGC-7901 and BGC-823 were increased significantly after treated with aspirin+chloroquine and aspirin+3-MA， while the distribution rate of SGC-7901 and BGC-823 cells at G1 phase were decreased significantly， with statistical significance （P＜0.05 or P＜0.01）. Compared with negative control group， there were no obvious DNA fragmentation fragments， apoptotic bodies and fragments of dense bright blue， while the number of migration cells were decreased significantly in SGC-7901 and BGC-823 cells after treated with aspirin （P＜0.001）； the number of autophagosome was increased significantly and the protein expression of LC3-Ⅱ was enhanced significantly （P＜0.05）. CONCLUSIONS： Aspirin can significantly inhibit the growth of SGC-7901 and BGC-823 cells， and arrest cell cycle in G1 phase， the mechanism of which may be associated with the activation of autophagy.
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OBJECTIVE： To study the effects of ivermectin on the migration and invasion of human gastric cancer cell lines BGC-823 and MGC-803 and its mechanism. METHODS： After treated with 0， 2.5， 5， 10， 20， 40 μmol/L ivermectin for 24 h， inhibitory rate of human gastric cancer cell lines BGC-823 and MGC-803 were detected by MTT assay. Effects of 5 μmol/L ivermectin and phosphate buffercontaining 0.67‰ dimethyl sulfoxide （control group） for 24 h on the migration and invasion of` gastric cancer cells BGC-823 and MGC-803 were observed by Transwell chamber invasion assay.Western blot assay was used to detect the protein expression of TGF-β1， TGF-βR， Smad2 and Smad3 in epithelial-mesenchymal transition （EMT） markers E-cadherin， N-cadherin， Vimentin， Snail and EMT transduction pathway TGF-β/smad of BGC-823 and MGC-803 cells after treated with 5， 10 μmol/L ivermectin and phosphate buffercontaining 0.67‰ dimethyl sulfoxide （control group） for 24 h. RESULTS： Ivermectin could inhibit the growth of BGC-823 and MGC-803， inhibitory rate of it was positively correlated with its concentration. Compared with control group， the number of migration and invasion BGC-823 and MGC-803 cells were decreased significantly after treated with 5 μmol/L ivermectin （P＜0.01 or P＜0.001）； the expression of E-cadherin protein was enhanced significantly in BGC-823 and MGC-803 cells after treated with 5 and 10 μmol/L ivermectin （P＜0.05 or P＜0.01 or P＜0.001）； the protein expression of N-cadherin， Vimentin， Snail， TGF-βR， Smad2 and Smad3 were decreased significantly （P＜0.05， P＜0.01 or P＜0.001）； protein expression of TGF-β1 was decreased significantly after treated with 10 μmol/L ivermectin （P＜0.05）. CONCLUSIONS： Ivermectin can significantly inhibit the migration and invasion of gastric cancer cells BGC-823 and MGC-803， and inhibiting the biological activity of EMT by reducing the expression of TGF-β/smad pathway is one of the mechanisms that inhibit the migration and invasion of gastric cancer cells.
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Objective:To investigate the influence of inhibitor of kappa B kinase ( IKK) inhibitor on carotid artery restenosis in rat by balloon catheter injury. Methods: A total of 32 male Wistar rats were randomly divided into control group, injured group and med?icine group. The balloon catheter injury was performed on left common carotid artery of rat by imitating the process of angioplasty. In?jured group and control group were injected saline, medicine group were injected IKK inhibitor. After 15 days, the process of neointi?mal and media hyperplasia was observed by HE staining and the expression level of NF?κB, IKK, inhibitor of nuclear factor kappa B ( IκB) was detected by Western blot, the content of IL?6, TNF?αwas detected by radioimmunoassay, and the content of MMP?1 was determined by MMP?1 kit. Results: Arterial neointima hyperplasia hapened in 3 groups. There was significantly difference between control group and medicine group and injured group in intimal area, the expression level of NF?κB, IKK, IκB, and the content of IL?6, TNF?α and MMP?1 ( P<0?05 or 0?01) . There was no significance in these indexes between control group and medicine group ( P>0?05) . Conclusion:IKK inhibitor can reduce the carotid artery restenosis after balloon injury by inhibiting inflammatory reaction induced by IKK?IκB?NF?κB pathway.