Emodin regulating excision repair cross-complementation group 1 through fibroblast growth factor receptor 2 signaling
Emodin regulating excision repair cross-complementation group 1 through fibroblast growth factor receptor 2 signaling
复制标题
大黄素通过成纤维细胞生长因子受体2信号传导调节切除修复交叉互补组1
DOI：
10.3748/wjg.v19.i16.2481
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发表时间：
2013-04-28
影响因子：
4.3
通讯作者：
Zou, Sheng-Quan
中科院分区：
文献类型：
--
作者：
Chen, Gang;Qiu, Hong;Zou, Sheng-Quan
目的：探讨大黄素逆转肝癌细胞对铂类药物耐药的分子机制。方法：将10 mU/L大黄素作用于人肝癌细胞株HepG2/奥沙利铂(OXA)后，计算细胞的抑制率(IR)、半数抑制浓度(IC50)和逆转指数(IC50)。将HepG2、HepG2/OxA、HepG2/OxA/T细胞分为对照组、OXA组、OXA+成纤维细胞生长因子7(FGF7)组和OXA+大黄素组，每组细胞的终浓度分别为5 ng/mL、10 mU g/mL和10 mU/L。单细胞凝胶电泳法检测成纤维细胞生长因子受体2(FGFR2)、磷酸化细胞外信号调节蛋白1/2(p-ERK1/2)和切除修复交叉互补基因1(ERCC1)的蛋白表达水平。结果：10 mU/L大黄素作用后，细胞内IC50由120.78 mU/L降至39.65 mU/L，逆转指数为3.0 5。与对照组相比，OXA组、OXA+Fgf7组、OXA+大黄素组的尾长和橄榄尾长均显著增加，差异有统计学意义(P<；0.01)。OXA+Fgf7组的尾长和橄榄尾长均低于OXA组，差异有统计学意义。与OXA+Fgf7组相比，OXA+大黄素组的尾长、Olive尾矩和尾部DNA百分比均显著增加，差异有统计学意义(P<；0.01)。与亲本细胞株HepG2相比，HepG2/OXA细胞中FGFR2、p-ERK1/2和ERCC1的表达水平显著升高，而在FGFR2 shRNA沉默的HepG2/OXA/T细胞中，这三种分子的表达均受到显著抑制。在检测的HepG2细胞中，OXA组和OXA+Fgf7组的FGFR2、p-ERK1/2和ERCC1的表达水平呈上升趋势。与OXA组和OXA+Fgf7组相比，OXA+大黄素组FGFR2、p-ERK1/2和ERCC1表达水平显著降低，差异有统计学意义。在转染FGFR2 shRNA的HepG2/OXA/T细胞中，FGFR2、p-ERK1/2和ERCC1的表达水平明显受到抑制，但OXA组、OXA+Fgf7组和OXA+大黄素组之间这些表达水平无显著差异。结论：大黄素通过增强OXA DNA损伤而显著逆转了OXA耐药，其分子机制与FGFR2/ERK1/2信号通路介导的抑制ERCC1表达有关。(C)2013年白石登。版权所有。
AIM: To investigate the molecular mechanisms underlying the reversal effect of emodin on platinum resistance in hepatocellular carcinoma.METHODS: After the addition of 10 mu mol/L emodin to HepG2/oxaliplatin (OXA) cells, the inhibition rate (IR), 50% inhibitory concentration (IC50) and reversal index (IC50 in experimental group/IC50 in control group) were calculated. For HepG2, HepG2/OXA, HepG2/OXA/T, each cell line was divided into a control group, OXA group, OXA + fibroblast growth factor 7 (FGF7) group and OXA + emodin group, and the final concentrations of FGF7, emodin and OXA in each group were 5 ng/mL, 10 mu g/mL and 10 mu mol/L, respectively. Single-cell gel electrophoresis was conducted to detect DNA damage, and the fibroblast growth factor receptor 2 (FGFR2), phosphorylated extracellular signal-regulated kinase 1/2 (p-ERK1/2) and excision repair cross-complementing gene 1 (ERCC1) protein expression levels in each group were examined by Western blotting.RESULTS: Compared with the IC50 of 120.78 mu mol/L in HepG2/OXA cells, the IC50 decreased to 39.65 mu mol/L after treatment with 10 mu mol/L emodin; thus, the reversal index was 3.05. Compared with the control group, the tail length and Olive tail length in the OXA group, OXA + FGF7 group and OXA + emodin group were significantly increased, and the differences were statistically significant (P < 0.01). The tail length and Olive tail length were lower in the OXA + FGF7 group than in the OXA group, and this difference was also statistically significant. Compared with the OXA + FGF7 group, the tail extent, the Olive tail moment and the percentage of tail DNA were significantly increased in the OXA + emodin group, and these differences were statistically significant (P < 0.01). In comparison with its parental cell line HepG2, the HepG2/OXA cells demonstrated significantly increased FGFR2, p-ERK1/2 and ERCC1 expression levels, whereas the expression of all three molecules was significantly inhibited in HepG2/OXA/T cells, in which FGFR2 was silenced by FGFR2 shRNA. In the examined HepG2 cells, the FGFR2, p-ERK1/2 and ERCC1 expression levels demonstrated increasing trends in the OXA group and OXA + FGF7 group. Compared with the OXA group and OXA + FGF7 group, the FGFR2, p-ERK1/2, and ERCC1 expression levels were significantly lower in the OXA + emodin group, and these differences were statistically significant. In the HepG2/OXA/T cell line that was transfected with FGFR2 shRNA, the FGFR2, p-ERK1/2 and ERCC1 expression levels were significantly inhibited, but there were no significant differences in these expression levels among the OXA, OXA + FGF7 and OXA + emodin groups.CONCLUSION: Emodin markedly reversed OXA resistance by enhancing OXA DNA damage in HepG2/OXA cells, and the molecular mechanism was related to the inhibitory effect on ERCC1 expression being mediated by the FGFR2/ERK1/2 signaling pathway. (C) 2013 Baishideng. All rights reserved.