目的 探究川陳皮素調(diào)節(jié)腎素血管緊張素系統(tǒng)（RAS）/蛋白激酶（RAF）/促分裂原活化蛋白激酶激酶（MEK）／細(xì)胞外信號(hào)調(diào)節(jié)激酶（ERK）信號(hào)通路對(duì)直腸癌細(xì)胞增殖、遷移和侵襲的影響。方法 用質(zhì)量濃度為5、10、20、40、80 μg·mL^-1的川陳皮素處理人直腸癌細(xì)胞（SW480），篩選合適的實(shí)驗(yàn)濃度；隨機(jī)將SW480細(xì)胞分為對(duì)照組，川陳皮素低、中、高質(zhì)量濃度（10、20、40 μg·mL^-1）組，川陳皮素40 μg·mL^-1+RAS激活劑（RASALl）組；檢測(cè)SW480細(xì)胞增殖、遷移、侵襲能力；Western blotting檢測(cè)蛋白表達(dá)；裸鼠成瘤實(shí)驗(yàn)檢測(cè)川陳皮素對(duì)直腸腫瘤的影響。結(jié)果 與對(duì)照組比較，川陳皮素低、中、高質(zhì)量濃度組5-乙炔基-2'-脫氧尿嘧啶核苷（Edu）陽(yáng)性率、細(xì)胞侵襲數(shù)、劃痕愈合率下降，細(xì)胞周期蛋白依賴(lài)性激酶1（CDK1）、基質(zhì)金屬蛋白酶-2（MMP-2）、基質(zhì)金屬蛋白酶-9（MMP-9）、RAS、RAF、MEK、ERK蛋白表達(dá)水平下降（P＜0.05）；與川陳皮素高質(zhì)量濃度組相比，RASALl組Edu陽(yáng)性率、細(xì)胞侵襲數(shù)、劃痕愈合率升高，CDK1、MMP-2、MMP-9、RAS、RAF、MEK、ERK蛋白表達(dá)水平升高（P＜0.05）。川陳皮素組移植瘤生長(zhǎng)速度比對(duì)照組緩慢，瘤體積減小，瘤質(zhì)量下降，RAS、RAF、MEK、ERK蛋白表達(dá)下調(diào)（P＜0.001）。結(jié)論 川陳皮素抑制RAS/RAF/MEK/ERK信號(hào)通路的激活，進(jìn)而抑制直腸癌細(xì)胞增殖、遷移和侵襲。

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 80 μg·mL^-1, and suitable experimental concentrations were screened. SW480 cells were randomly divided into control group, nobiletin low-, mid-, and high-doses (10, 20, 40 μg·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 40 μg·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.

R285.5

湖北科技學(xué)院2020年五官醫(yī)學(xué)院專(zhuān)項(xiàng)科研基金項(xiàng)目(2020WG07)