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Objective To evaluate the effect of silencing ACAT1 gene on colon cancer cells proliferation,migration,invasion and colon cancer development by using the small interference RNA (siRNA) in colon cancer cell line HT-29.Methods Acyl coenzyme A cholesterol acyltransferase 1 (ACAT1) gene was silenced in HT-29 cell lines using Hiperfect transfection reagent.The expression level of ACAT1 was detected by real time PCR.CFSE and transwell assays were used to evaluate the effect of ACAT1 gene interfering on cells proliferation,mi gration and invasion.Result ACAT1 mRNA expression decreased obviously after siRNA interference.Compared with pre-transfection,proliferation,migration and invasion of colon cancer cells have been significantly inhibited (P ＜ 0.05).Conclusion ACAT1 gene interference reduced proliferation,migration and of invasion of HT29 cells,which provide a new potential target for colon cancer treatment.
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OBJECTIVE:To investigate the inhibitory effect of ligustrazine on the formation of macrophage-derived foam cells and its possible mechanism. METHODS:Human acute mononuclear cells (THP-1) were incubated with 160 nmol/L phorbol ester (PMA) for 24 h to differentiate into macrophages;and the macrophages were incubated with oxidized low-density lipoprotein (ox-LDL)culture solution containing 80 mg/L for 24 h to differentiate into macrophage-derived foam cells. And then the cells were randomly divided into blank control group(ox-LDL),model(AngⅡ)group,positive control(valsartan)group,and ligustrazine low,medium and high concentration groups(the mass concentration were 0.025,0.05 and 0.1 g/L). After all cells were respective-ly incubated with 80 mg/L ox-LDL culture solution for 48 h,oil red O staining was adopted to observe the transformation rate of foam cells,enzyme chemical method was used to determine the content of cholesterol,and real-time quantitative polymerase chain (RT-PCR)and Western blot were conducted to detect expression levels of Acyl-coenzyme A cholesterol acyltransferase-1(ACAT-1) mRNA and its protein. RESULTS:Compared with blank control group,the transformation rate of foam cells and content of choles-terol in model group were increased,and the expression levels of ACAT-1 mRNA and its protein were obviously strengthened,with significant differences(P<0.01 or P<0.05). Compared with model group,the transformation rate of foam cells and content of cho-lesterol in positive control group(valsartan)and ligustrazine low,medium and high concentration groups were decreased,and the expression levels of ACAT-1 mRNA and its protein were obviously weakened,with significant differences (P<0.01 or P<0.05). CONCLUSIONS:Ligustrazine can inhibit the macrophages differentiating into foam cells,by a mechanism that may be related to inhibiting expression of ACAT-1,and reducing content of cholesterol to reduce formation of foam cells.
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Objective: To study the effects of total flavonoids from Dracocephalum moldovica on the formation of mouse peritoneal macrophage-derived foam cells and their possible mechanisms. Methods: The mouse peritoneal macrophages were induced with oxidized low density lipoprotein (Ox-LDL) to establish foam cell model, and were intervened with different concentration of total flavonoids from D. moldovica. The foam cells were observed by oil red O staining and the contents of cholesterol ester in cells were detected by fluorescence spectrophotometric method. The expression levels of acyl-coenzyme A, cholesterol acyltransferase-1 (ACAT-1) mRNA in cells and scavenger receptor A (SR-A) on cells were determined using real-time quantitative PCR (RT-qPCR) and flow cytometry, respectively. Results: The Ox-LDL-induced formation of foam cells, accumulation of cholesterol ester in cells, and the expression levels of ACAT-1 mRNA in cells and SR-A on cells were significantly inhibited by total flavonoids from D. moldovica, which showed a tendency toward a dose-dependent manner. Conclusion: These data suggest that total flavonoids from D. moldovica could inhibit the formation of macrophage-derived foam cells, which is probably one of the underlying mechanisms of the total flavonoids for clinical treatment of atherosclerosis.
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Objective To investigate the effects of advanced glycosylation end-products (AGEs) on the expression of acyl coenzyme A: cholesterol acyltransferase-1 (ACAT-1) in cultured THP-1 macrophages. Methods THP-1 cells were differentiated into macrophages after cultured in RPMI1640 media containing 0.1 ?mol/L PMA for 72 h. THP-1 macrophages were then exposed to AGE-modified bovine serum albumin (AGE-BSA, at concentration of 50, 100, 200 mg/L) for 24 h or to 200 mg/L AGE-BSA for 0, 12, 24, 36 h. Expression of ACAT-1 mRNA and protein in THP-1 macrophages was measured by RT-PCR and Western blot, respectively. Results After induced by 0.1 ?mol/L PMA, THP-1 cells stopped proliferation and differentiated into macrophages. Treatment of THP-1 macrophages with AGE-BSA resulted in an increase in the mRNA and protein levels for ACAT-1 in a dose-and time-dependent manner. Conclusion AGEs upregulate the expression levels of mRNA and protein for ACAT-1 in cultured THP-1 macrophages, which might be partly involved in the atherogenesis in diabetic patients.
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Objective To investigate the effects of valsartan on the expression of acyl-coenzyme A: cholesterol acyltransferase-1(ACAT-1) and peroxisome proliferator activated receptor-gamma(PPAR-?) in atherosclerotic plaques on rabbit aortic wall.Methods Twenty-four male Japanese white rabbits were randomly assigned into three groups(8 each): control group,valsartan group and high cholesterol feeding group.All rabbits were fed according to the experimental protocol for 12 weeks.Blood samples were taken from vein for measurement of serum lipids.The ratio of intima/media thickness of the aorta was measured.ACAT-1 mRNA/protein and PPAR-? mRNA/protein were determined by reverse transcription polymerase chain reaction(RT-PCR) and Western blotting,respectively.Results After 12 weeks,the levels of serum total cholesterol(TC),triglyeride(TG) and low density lipoprotein-cholesterol(LDL-C) in valsartan group and cholesterol group were significantly higher than those in control group(P0.05).The intima thickness and the ratio of intima/media in carotid arteries in cholesterol group were significantly higher than those in control group and valsartan group(P