摘要
目的探讨吲哚-3-甲醇(I3C)对神经母细胞瘤(NB)细胞增殖与迁移能力的影响及其机制。方法采用CCK8实验检测终浓度为0、50、100、150、200、300、500μmol/L的I3C对SH-SY5Y细胞活力的影响,并计算I3C对SH-SY5Y细胞的半数致死浓度(IC50值),作为后续实验中I3C浓度设定的参考值。以IC50值为依据,SH-SY5Y细胞中分别加入终浓度为0、30、60、90μmol/L的I3C(分别为A、B、C、D组),通过划痕实验、实时荧光定量PCR(RT-qPCR)技术、Western Blot实验分别检测各组细胞的迁移能力及细胞中LSD1 mRNA及LSD1、H3K4me1、H3K4me2蛋白的相对表达量。结果CCK8实验结果显示,随着I3C浓度的递增,SH-SY5Y细胞活力明显下降(F=1823.19,P<0.05),计算得出I3C作用SH-SY5Y细胞48 h的IC50值为121.00μmol/L;划痕实验结果显示,A~D组SH-SY5Y细胞的迁移能力随着I3C浓度的增高而明显下降(F=388.47,P<0.05),各组间比较差异具有显著性(t=8.86~33.43,P<0.05);RT-qPCR技术及Western Blot实验检测结果显示,SH-SY5Y细胞中LSD1 mRNA及蛋白相对表达量均随I3C浓度的增高而明显降低(F=27.67、522.41,P<0.05),各组间比较差异均有显著性(t=4.16~56.28,P<0.05),A~D组SH-SY5Y细胞中H3K4me1、H3K4me2蛋白相对表达量随着I3C浓度的增高逐渐升高(F=36.53、54.60,P<0.05)。结论I3C具有明显抑制SH-SY5Y细胞增殖与迁移的能力,其抑制作用可能是通过调节LSD1的表达实现的。
Objective To investigate the effect of indole-3-carbinol(I3C)on the proliferation and migration of neuroblastoma(NB)cells and its mechanism.Methods CCK8 assay was used to investigate the effect of I3C with a final concentration of 0,50,100,150,200,300,and 500μmol/L on the viability of SH-SY5Y cells,and the median lethal concentration(IC50)value of I3C on SH-SY5Y cells was calculated and was used as the reference value for determining I3C concentration in subsequent experiments.Based on the IC50 value,SH-SY5Y cells were treated with I3C at a final concentration of 0,30,60,and 90μmol/L(groups A,B,C,and D,respectively),and wound healing assay,quantitative real-time PCR,and Western Blot were used to measure the migration ability of cells,the relative mRNA expression level of LSD1,and the relative protein expression levels of LSD1,H3K4me1,and H3K4me2.Results CCK8 assay showed that the viability of SH-SY5Y cells decreased significantly with the increase in I3C concentration(F=1823.19,P<0.05),and the IC50 value of I3C treatment for 48 hours was calculated to be 121.00μmol/L for SH-SY5Y cells.Wound healing assay showed that the migration ability of SH-SY5Y cells in groups A-D decreased significantly with the increase in I3C concentration(F=388.47,P<0.05),and there were significant differences between groups(t=8.86-33.43,P<0.05).Quantitative real-time PCR and Western Blot showed that the relative mRNA and protein expression levels of LSD1 in SH-SY5Y cells significantly decreased with the increase in I3C concentration(F=27.67,522.41,P<0.05),with significant differences between groups(t=4.16-56.28,P<0.05),and the relative protein expression levels of H3K4me1 and H3K4me2 in SH-SY5Y cells in groups A-D gradually increased with the increase in I3C concentration(F=36.53,54.60,P<0.05).Conclusion I3C significantly inhibits the proliferation and migration of SH-SY5Y cells,possibly by regulating the expression of LSD1.
作者
周娜
丁伟
宋金莲
王军
郑文祥
侯琳
ZHOU Na;DING Wei;SONG Jinlian;WANG Jun;ZHENG Wenxiang;HOU Lin(Department of Biochemistry and Molecular Biology, School of Medicine, Qingdao University, Qingdao 266021, China)
出处
《精准医学杂志》
2021年第2期132-135,共4页
Journal of Precision Medicine
基金
山东省重点研发项目(2019GSF107025)
青岛创业创新领军人才计划(13-CX-3)。
