Xu Lei, Wu Huini, Jiang Chao, Wang Hongcai, Gao Bei, Yan Shumei, Qi Yushu, Zhou Shufeng
Maternal and Child Health Hospital of Zibo City, Shandong 255022, China.
Department of Biological Sciences, University of Illinois at Chicago, Chicago, IL 60607, USA.
Discov Med. 2016 Nov;22(122):297-309.
Aberrant epidermal growth factor receptor (EGFR) is involved in a variety of cancers and its inhibitors have been studied for over a decade. We aim to investigate the effects of dacomitinib, a second generation pan-EGFR inhibitor, on ovarian cancer cells.
By immunohistochemistry, we studied the clinical significance of EGFR expression in epithelial ovarian cancer (EOC). The correlations between EGFR expression and the clinicopathological variables of patients with EOC were assessed using Pearson's X2 test. Kaplan-Meier analysis was used to compare the postoperative survival between groups of patients with EOC with varying levels of EGFR expression. MTT, caspase assay, cell apoptosis analysis, autophagy analysis, cell cycle analysis, and western blotting were used to investigate various effects of dacomitinib in cell proliferation, apoptosis, and associated molecular pathways.
High expression of EGFR was found to be associated with poor prognosis of patients with EOC. EGFR, P-AKT, and P-ERK were inhibited after treatment of dacomitinib in both SKOV3 and OV4 cells. Activations of caspase activities, apoptosis, and autophagy were also observed and confirmed by western blot: caspase 9, LC3, and Bax levels were elevated, while Bcl-2 and Bcl-xl were down-regulated. The percentage of cancer cells in the S and G2 phases of the cell cycle significantly decreased after treatment. Cdk1 and Cdk2 protein levels declined after dacomitinib treatment; epithelial-mesenchymal transition (EMT) was inhibited, which was confirmed by observing E-cadherin, N-cadherin, and slug inhibition. Additionally, dacomitinib significantly increased chemotherapy sensitivity in chemotherapeutic resistant ovarian cell lines, C13 and 2780CP.
Our data showed that increased expression of EGFR is associated with poor prognosis of patients with EOC and dacomitinib may act as a novel, useful chemotherapy drug. Further studies are warranted.
异常的表皮生长因子受体（EGFR）参与多种癌症，其抑制剂已被研究了十多年。我们旨在研究第二代泛EGFR抑制剂达可替尼对卵巢癌细胞的影响。
通过免疫组织化学，我们研究了EGFR表达在上皮性卵巢癌（EOC）中的临床意义。使用Pearson卡方检验评估EGFR表达与EOC患者临床病理变量之间的相关性。采用Kaplan-Meier分析比较不同EGFR表达水平的EOC患者组术后生存率。采用MTT法、半胱天冬酶检测、细胞凋亡分析、自噬分析、细胞周期分析和蛋白质印迹法研究达可替尼对细胞增殖、凋亡及相关分子途径的各种影响。
发现EGFR高表达与EOC患者预后不良相关。在SKOV3和OV4细胞中，达可替尼治疗后EGFR、P-AKT和P-ERK均受到抑制。半胱天冬酶活性、细胞凋亡和自噬的激活也通过蛋白质印迹得到观察和证实：半胱天冬酶9、LC3和Bax水平升高，而Bcl-2和Bcl-xl下调。细胞周期S期和G2期癌细胞百分比在治疗后显著降低。达可替尼治疗后Cdk1和Cdk2蛋白水平下降；上皮-间质转化（EMT）受到抑制，通过观察E-钙黏蛋白、N-钙黏蛋白和蛞蝓抑制得到证实。此外，达可替尼显著提高了化疗耐药卵巢细胞系C13和2780CP的化疗敏感性。
我们的数据表明，EGFR表达增加与EOC患者预后不良相关，达可替尼可能是一种新型、有效的化疗药物。有必要进行进一步研究。
