川陈皮素调节RAS/RAF/MEK/ERK信号通路对直肠癌细胞增殖、迁移和侵袭的影响

Effects of nobiletin on proliferation,migration,and invasion of rectal cancer cells by regulating RAS/RAF/MEK/ERK signaling pathway

目的探究川陈皮素调节肾素血管紧张素系统(RAS)/蛋白激酶(RAF)/促分裂原活化蛋白激酶激酶(MEK)/细胞外信号调节激酶(ERK)信号通路对直肠癌细胞增殖、迁移和侵袭的影响。方法用质量浓度为5、10、20、40、80μg·mL^(−1)的川陈皮素处理人直肠癌细胞(SW480),筛选合适的实验浓度;随机将SW480细胞分为对照组,川陈皮素低、中、高质量浓度(10、20、40μg·mL^(−1))组,川陈皮素40μg·mL^(−1)+RAS激活剂(RASALl)组;检测SW480细胞增殖、迁移、侵袭能力;Western blotting检测蛋白表达;裸鼠成瘤实验检测川陈皮素对直肠肿瘤的影响。结果与对照组比较,川陈皮素低、中、高质量浓度组5-乙炔基-2'-脱氧尿嘧啶核苷(Edu)阳性率、细胞侵袭数、划痕愈合率下降,细胞周期蛋白依赖性激酶1(CDK1)、基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)、RAS、RAF、MEK、ERK蛋白表达水平下降(P<0.05);与川陈皮素高质量浓度组相比,RASALl组Edu阳性率、细胞侵袭数、划痕愈合率升高,CDK1、MMP-2、MMP-9、RAS、RAF、MEK、ERK蛋白表达水平升高(P<0.05)。川陈皮素组移植瘤生长速度比对照组缓慢,瘤体积减小,瘤质量下降,RAS、RAF、MEK、ERK蛋白表达下调(P<0.001)。结论川陈皮素抑制RAS/RAF/MEK/ERK信号通路的激活,进而抑制直肠癌细胞增殖、迁移和侵袭。

Objective To investigate the effects of nobiletin on the proliferation,migration,and invasion of rectal cancer cells by regulating the renin-angiotensin system(RAS)/RAF/mitogen-activated protein kinase(MEK)/extracellular regulated protein kinase(ERK)signaling pathway.Methods Human rectal cancer cells(SW480)were treated with nobiletin at concentrations of 5,10,20,40,and 80μg·mL^(−1),and suitable experimental concentrations were screened.SW480 cells were randomly divided into control group,nobiletin low-,mid-,and high-doses(10,20,40μg·mL^(−1))groups,and a high dose nobiletin+RAS activator group(RASALl group).The proliferation,migration,and invasion ability of SW480 cells were detected.Western blotting was applied to detect protein expression.Nude mice were used to detect the effect of nobiletin on rectal tumors.Results By screening the concentrations,concentrations of 10,20,and 40μg·mL^(−1) of nobiletin were selected for the experiment.Compared with the control group,the Edu positive rate,cell invasion number,and scratch healing rate decreased in the nobiletin groups,while the expression levels of cyclindependent kinase 1(CDK1),matrix metalloproteinase-2(MMP-2),matrix metalloproteinase-9(MMP-9),RAS,RAF,MEK,and ERK decreased(P<0.05).Compared with nobiletin high dose group,the Edu positive rate,cell invasion number,and scratch healing rate increased in the nobiletin+RASALl group,while the expression levels of CDK1,MMP-2,MMP-9,RAS,RAF,MEK,and ERK(P<0.05).The growth rate of transplanted tumors in the nobiletin groups was slower than that in the control group,the tumor volume decreased,the quality decreased,and the expression of RAS,RAF,MEK,and ERK proteins was down regulated(P<0.05).Conclusion Nobiletin inhibits the activation of the RAS/RAF/MEK/ERK signaling pathway,thereby inhibiting the proliferation,migration,and invasion of rectal cancer cells.