基于miR-181/NF-κB通路的辛伐他汀对结直肠癌小鼠血液高凝状态的改善研究

Improvement of Blood Hypercoagulability in Mice with Colorectal Cancer by Simvastatin Based on miR-181/NF-κB Pathway

目的:基于微小RNA(miR)-181/核因子κB(NF-κB)通路,探讨辛伐他汀对结直肠癌小鼠肿瘤生长、血液高凝状态的影响。方法:选取BALB/C裸鼠,通过皮下注射人结直肠癌细胞系SW480的方法构建结直肠癌小鼠模型,将30只模型小鼠随机分为对照组、辛伐他汀低和高剂量组。辛伐他汀低、高剂量组小鼠分别腹腔注射辛伐他汀50、100 mg/kg, 1日1次,第28日分析小鼠肿瘤生长、血管生成和凝血情况。通过定量反转录聚合酶链反应、蛋白质印迹法检测肿瘤组织中miR-181、NF-κB抑制蛋白α(IκBα)以及NF-κB mRNA和蛋白表达水平。结果:与对照组比较,辛伐他汀低、高剂量组小鼠的肿瘤体积和重量,细胞周期蛋白D1(CyclinD1)、Ki67和B细胞淋巴瘤2(Bcl-2)蛋白水平,微血管密度、血管内皮生长因子(VEGF)水平和全血黏度,IκBα、NF-κB mRNA和蛋白表达水平均显著降低;而Bcl-2相关X蛋白(Bax)、miR-181水平显著升高,血浆凝血酶原时间(PT)、活化部分凝血活酶时间(APTT)显著延长,上述差异均有统计学意义(P<0.05)。辛伐他汀高剂量组小鼠的肿瘤体积和重量、微血管密度显著低于辛伐他汀低剂量组[(171.74±19.33)mm^(3) vs.(226.52±26.38)mm^(3),(0.21±0.03)g vs.(0.28±0.04)g,(9.77±1.85)%vs.(16.39±2.11)%],CyclinD1、Ki67和Bcl-2蛋白水平,VEGF水平、全血黏度,IκBα、NF-κB mRNA和蛋白表达水平显著低于辛伐他汀低剂量组;Bax蛋白、miR-181水平显著高于辛伐他汀低剂量组,PT、APTT显著长于辛伐他汀低剂量组,上述差异均有统计学意义(P<0.05)。结论:辛伐他汀可能通过调控miR-181/NF-κB途径抑制结直肠癌模型小鼠的肿瘤生长和血管生成,并缓解血液高凝状态。

OBJECTIVE:To explore the effects of simvastatin on tumor growth,blood coagulation in mice with colorectal cancer based on microRNA(miR)-181/nuclear factor(NF)-κB pathway.METHODS:The colorectal cancer mice model was established by subcutaneous injection of human colorectal cancer cell line SW480,and 30 model mice were randomly divided into the control group,simvastatin low-dose group and simvastatin high-dose group.The simvastatin low-dose group and simvastatin high-dose group were intraperitoneally injected with 50 mg/kg and 100 mg/kg simvastatin,qd.On the 28th day,tumor growth,angiogenesis,coagulation indicators were analyzed.The expression levels of miR-181,IκBαand NF-κB mRNA and protein in tumor tissues were detected by quantitative reverse transcription polymerase chain reaction and western blotting.RESULTS:Compared with the control group,the tumor volume,mass,CyclinD1 protein,Ki67 protein,Bcl-2 protein,microvessel density,vascular endothelial growth factor(VEGF)levels,whole blood viscosity,IκBαand NF-κB mRNA and protein expression levels of the simvastatin low-dose group and simvastatin high-dose group decreased significantly,while Bax protein and miR-181 levels increased,plasma prothrombin time(PT),activated partial thromboplastin time(APTT)and miR-181 were prolonged significantly,with statistically significant differences(P<0.05).The tumor volume,mass and microvascular density in simvastatin high-dose group were significantly lower than those in simvastatin low-dose group[(171.74±19.33)mm^(3) vs.(226.52±26.38)mm^(3),(0.21±0.03)g vs.(0.28±0.04)g,(9.77±1.85)%vs.(16.39±2.11)%],CyclinD1,Ki67 and Bcl-2 protein levels,VEGF levels,whole blood viscosity,expression levels of IκBαand NF-κB mRNA and protein were significantly lower than those in simvastatin low-dose group;the levels of Bax protein and miR-181 were significantly higher than those of simvastatin low-dose group,PT and APTT were significantly longer than those of simvastatin low-dose group,the differences were statistically significant(P<0.05).CONCLUSIONS:Simvastatin may inhibit tumor growth and angiogenesis in colorectal cancer model mice by regulating the miR-181/NF-κB pathway,and alleviate blood hypercoagulability.