The fibroblast growth factor receptor 2-mediated extracellular signal-regulated kinase 1/2 signaling pathway plays is important in regulating excision repair cross-complementary gene 1 expression in hepatocellular carcinoma
The fibroblast growth factor receptor 2-mediated extracellular signal-regulated kinase 1/2 signaling pathway plays is important in regulating excision repair cross-complementary gene 1 expression in hepatocellular carcinoma
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DOI：
10.3892/br.2013.96
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发表时间：
2013-07-01
影响因子：
2.3
通讯作者：
Zou, Shengquan
中科院分区：
文献类型：
--
作者：
Chen, Gang;Qiu, Hong;Zou, Shengquan
切除修复交叉互补基因1（ERCC 1）是成纤维细胞生长因子受体2（FGFR 2）的下游调控靶点，其作用机制尚未阐明。FGFR 2下游的级联反应包括PKC、Ras、Raf、MEK、ERK、JAK、STAT和PI 3 K途径。ERCC 1被认为是细胞外信号调节激酶（ERK）1/2的密切相关的下游靶基因，因为ERCC 1 mRNA和蛋白水平可被ERK抑制剂U 0126抑制。据推测，与成纤维细胞生长因子7（FGF 7）特异性结合的FGFR 2可能通过ERK信号通路调节ERCC 1基因表达。本研究旨在探讨FGFR 2对肝癌耐药细胞株ERCC 1基因表达的调控作用与p-ERK 1/2信号通路之间的关系。将Bek shRNA转染对数生长期的耐药细胞系HepG 2/OXA及其亲本细胞系HepG 2。转染和未转染的HepG 2和HepG 2/OXA细胞用FGF 7刺激，Western blot分析FGFR 2、p-ERK 1/2和ERCC 1蛋白表达的变化。转染后，观察到HepG2.T和HepG2.OXA. T细胞在含有嘌呤霉素的筛选培养基中稳定生长。Western blot分析显示，转染后的HepG2.T和HepG 2/OXA/T细胞中FGFR 2、p-ERK 1/2和ERCC 1蛋白表达水平均显著低于未转染细胞。HepG 2/OXA细胞中FGFR 2、p-ERK 1/2和ERCC 1的表达较亲本HepG 2细胞显著增加。FGF 7刺激后，HepG 2细胞中FGFR 2、p-ERK 1/2和ERCC 1的表达增加，其中p-ERK 1.2和ERCC 1的表达在HepG 2和HepG 2/OXA细胞中差异有统计学意义。Bek shRNA转染HepG 2. T和HepG 2/OXA/T细胞后蛋白水平无差异。结论：肝癌细胞中FGFR 2介导的ERK 1/2信号通路在ERCC 1表达调控中起重要作用。
Excision repair cross-complementary gene 1 (ERCC1) is a downstream regulatory target of fibroblast growth factor receptor 2 (FGFR2); however, the mechanism of its action has not been elucidated. The cascades downstream of FGFR2 include the PKC, Ras.Raf.MEK.ERK, JAK.STAT and PI3K pathways. ERCC1 is considered to be a closely related downstream target gene of extracellular signal-regulated kinase (ERK) 1/2, since ERCC1 mRNA and protein levels may be inhibited by the ERK inhibitor U0126. It was hypothesized that FGFR2, which specifically binds with fibroblast growth factor 7 (FGF7), may regulate ERCC1 gene expression through the ERK signaling pathway. The aim of the present study was to explore the association between the regulatory effect of FGFR2 on ERCC1 gene expression and the p-ERK1/2 signaling pathway in a drug-resistant hepatocellular carcinoma (HCC) cell line. The drug-resistant cell line HepG2/OXA and its parental cell line HepG2 were transfected with Bek shRNA in the logarithmic growth phase. Transfected and untransfected HepG2 and HepG2/OXA cells were then stimulated with FGF7 and changes in the protein expression of FGFR2, p-ERK1/2 and ERCC1 was detected with western blot analysis. Following transfection, HepG2.T and HepG2.OXA.T cells were observed to grow stably in a screening medium containing puromycin. The western blot analysis demonstrated a significant decrease in the protein expressions of FGFR2, p-ERK1/2 and ERCC1 in HepG2.T and HepG2/OXA/T cells as compared to untransfected cells. Expression of FGFR2, p-ERK1/2 and ERCC1 in HepG2/OXA cells was significantly increased compared to the parental HepG2 cells. Following stimulation with FGF7, the expression of FGFR2, p-ERK1/2 and ERCC1 was increased, with significant differences between HepG2 and HepG2/OXA cells in the expression of p-ERK1.2 and ERCC1. No differences were detected in the protein levels following Bek shRNA transfection in HepG2.T and HepG2/OXA/T cells. In conclusion, the FGFR2-mediated ERK1/2 signaling pathway in HCC cells plays an important role in the regulation of ERCC1 expression.