姜黄素对人胰腺癌细胞增殖抑制作用及对Wnt信号通路的影响

Effects of Curcumin on Proliferation Inhibition of Human Pancreatic Cancer Cell and the Influence on Wnt Signaling Pathway

目的:探究姜黄素对人胰腺癌SW1990细胞增殖抑制作用及对Wnt信号通路的影响。方法:低/中/高剂量观察组人胰腺癌SW1990细胞分别以终浓度为20、50、100μmol/L姜黄素处理,100μL/孔,对照组细胞则以等体积生理盐水处理,分别干预24、48、72 h。以噻唑蓝(MTT)法及Annexin-V/FITC双染法分别检测人胰腺癌细胞SW1990增殖及凋亡情况,反转录聚合酶链式反应(RT-PCR)法及细胞爬片免疫细胞化学技术检测人胰腺癌细胞SW1990β-链蛋白(β-Catenin)mRNA及β-Catenin蛋白表达情况。结果:干预72 h后低/中/高剂量观察组人胰腺癌SW1990细胞增殖抑制率(%)分别为(4.59±1.32)、(40.01±3.87)、(60.98±5.97);对照组及低/中/高剂量观察组人胰腺癌SW1990细胞凋亡率(%)(8.14±1.25)、(13.04±1.03)、(27.15±1.62)、(59.21±1.46);β-Catenin蛋白免疫细胞化学(ICC)评分(分)分别为(5.78±0.56)、(4.42±0.61)、(2.57±0.58)、(1.38±0.63)。观察组凋亡率显著高于对照组,β-Catenin mRNA相对表达量及β-Catenin蛋白ICC评分显著低于对照组(P<0.05),且增殖抑制率、凋亡率随姜黄素作用浓度增加逐渐升高(P<0.05),增殖抑制率亦随干预时间延长逐渐升高(P<0.05);β-Catenin mRNA相对表达量及β-Catenin蛋白ICC评分随姜黄素作用浓度增加逐渐降低(P<0.05)。结论:姜黄素可有效下调人胰腺癌细胞SW1990β-Catenin基因表达,抑制Wnt信号通路活化,具有显著的增殖抑制作用,且具有明显的时间-剂量依赖性。

Objective：To explore the proliferation inhibition effect of curcumin on human pancreatic cancer SW1990 cell and the inffluenee on Wnt signaling pathway-. Methods：The human pancreatic cancer cells SW1990 in experimental-L/M/H group were treated with final concentration of 20, 50, 100 Ixmol/L eureumin, 100 uL/hole, and cells in control group were treated with iso-volumetric saline. Each group was intervened for 24, 48, 72 h respectively. The proliferation and apoptosis were detected by meth-yl thiazolyl tetrazolium （MTT） method and AnnexinV/FITC double staining respectively, expression d -Catenin mRNA and β- Catenin mRNA protein in human pancreatic cancer SW1990 cell were detected by reverse transcription PCtl （tlT-PCtl） method and cell climbing imnmnoeytoehenlistry technique respectively. Results：72 h after intervened, the proliferation inhibition rates （ % ） of human pancreatic cancer SW1990 cell in experimental-L/M/H group were 4. 59 ± 1.32, 40. 01 ± 3.87, 60. 98 ± 5.97 re-spectively; Apoptosis rates （%） in control group and experimental-L/M/H group were 8.14 ± 1.25, β.04 ±1.03, 27. 15 ±1.62, 59. 21 ±1.46 respectively; β-Catenin protein immunocytochemistry （ICC） score （score） were 5.78 ± 0. 56, 4.42 _＋ 0. 61, 2. 57 ± 0. 58, 1.38 ± 0. 63 respectively. The apoptosis rate in experimental groups were higher than that in control, and β-Catenin mRNA relative expression and β-Catenin protein ICC score were lower than those in control group （ P 〈 0. 05 ） , and the prolifera- tion inhibition rate, apoptosis rate in experimental groups increased gradually with the increase of eureumin action concentration （P〈0.05）. Proliferation inhibition also increased gradually with the lengthen of action time （ P 〈 0. 05 ） ; the β-Catenin mRNA rela- tive expression and β-Catenin protein ICC score decreased gradually with the increase of eureumin action concentration （P 〈 0. 05 ）. Conclusion：Cureumin can down-regulate the expression of β-Catenin in human pancreatic cancer SW1990 cell effectively, inhibit the Wnt signaling pathway excitation, and has significant proliferation inhibition effect, which has an obvious time-dose de-pendence.