Department of Biology, Faculty of Arts and Science, Uludag University, Bursa, Turkey.
Department of Clinical Biochemistry, Faculty of Medicine, Istinye University, Istanbul, Turkey.
Biochim Biophys Acta Gen Subj. 2019 May;1863(5):903-916. doi: 10.1016/j.bbagen.2019.02.014. Epub 2019 Feb 27.
It has been known epidermal growth factor receptor (EGFR) frequently overexpressed in cervical cancer. High levels of EGFR expression in their tumors leads to a poor prognosis and inhibition frequently induces autophagy in cancer cells. This study aimed to investigate whether EGFR inhibition by canertinib induces autophagy and this induction influence the effect of Palladium (Pd) (II) complex and 5-fluorouracil (5-FU) especially in nontoxic doses.
Cytotoxicity was evaluated by using SRB assay. Apoptosis, autophagy, and EGFR key markers were determined by flow cytometry, fluorescence staining, and immunoblotting. Colony formation, invasion, and wound healing assays were performed to investigate cell proliferation, invasion, and migration, respectively.
Blocking EGFR by the pan-ErbB tyrosine kinase inhibitor canertinib inhibited cell growth of HeLa cervical cancer cells in combination with Pd(II) complex and 5-FU. Combination of canertinib and Pd(II) complex promotes autophagy and apoptosis of HeLa cancer cells via blockade of the PI3K/AKT and MAPK/ERK pathway, which leads to cervical cancer cell death. ROS accumulation and DNA damage were increased after combinatorial treatment which causes depolarization of the mitochondrial inner membrane and leads to apoptotic cell death. Canertinib combined with Pd(II) complex leads to inhibition of migration and invasion.
Inhibition of EGFR signaling by canertinib in combination with Pd(II) complex promotes apoptosis and autophagy via blockade of the PI3K/AKT and MAPK/ERK.
The cytotoxic activity of Pd(II) complex and 5-FU on HeLa cells is mediated by EGFR inhibition and autophagy induction, leading to activation of mitochondrial apoptotic cell death.
表皮生长因子受体（EGFR）在宫颈癌中经常过度表达。其肿瘤中 EGFR 表达水平高导致预后不良，抑制 EGFR 常诱导癌细胞自噬。本研究旨在探讨卡那替尼抑制 EGFR 是否诱导自噬，以及这种诱导是否会影响钯（II）复合物和 5-氟尿嘧啶（5-FU）的作用，尤其是在非毒性剂量下。
采用 SRB 法评估细胞毒性。通过流式细胞术、荧光染色和免疫印迹法测定细胞凋亡、自噬和 EGFR 关键标志物。通过集落形成、侵袭和划痕愈合实验分别研究细胞增殖、侵袭和迁移。
泛 ErbB 酪氨酸激酶抑制剂卡那替尼阻断 EGFR 与 Pd(II) 复合物和 5-FU 联合抑制 HeLa 宫颈癌细胞生长。卡那替尼与 Pd(II) 复合物联合通过阻断 PI3K/AKT 和 MAPK/ERK 通路促进 HeLa 癌细胞自噬和凋亡，导致宫颈癌细胞死亡。联合治疗后 ROS 积累和 DNA 损伤增加，导致线粒体内膜去极化和凋亡细胞死亡。卡那替尼联合 Pd(II) 复合物导致迁移和侵袭抑制。
卡那替尼抑制 EGFR 信号通路与 Pd(II) 复合物联合通过阻断 PI3K/AKT 和 MAPK/ERK 促进凋亡和自噬。
Pd(II) 复合物和 5-FU 对 HeLa 细胞的细胞毒性活性是通过 EGFR 抑制和自噬诱导介导的，导致线粒体凋亡细胞死亡的激活。
