Zhu Yu-Sheng, Zhou Si-Rui, Zhang Hui-Hui, Wang Tong, Chen Xiao-Dong
Faculty of Life Sciences and Medicine, Northwest University, Xi'an 710069, Shaanxi Province, China.
First Affiliated Hospital of Northwest University, Northwest University, Xi'an 710069, Shaanxi Province, China.
Int J Ophthalmol. 2024 Jun 18;17(6):1018-1027. doi: 10.18240/ijo.2024.06.05. eCollection 2024.
To explore the effect of epidermal growth factor receptor (EGFR) inhibition by erlotinib and EGFR siRNA on epidermal growth factor (EGF)-induced activation of retinal pigment epithelium (RPE) cells.
Human RPE cell line (ARPE-19 cells) was activated by 100 ng/mL EGF. Erlotinib and EGFR siRNA were used to intervene EGF treatment. Cellular viability, proliferation, and migration were detected by methyl thiazolyl tetrazolium (MTT) assay, bromodeoxyuridine (BrdU) staining assay and wound healing assay, respectively. EGFR/protein kinase B (AKT) pathway proteins and N-cadherin, α-smooth muscle actin (α-SMA), and vimentin were tested by Western blot assay. EGFR was also determined by immunofluorescence staining.
EGF treatment for 24h induced a significant increase of ARPE-19 cells' viability, proliferation and migration, phosphorylation of EGFR/AKT proteins, and decreased total EGFR expression. Erlotinib suppressed ARPE-19 cells' viability, proliferation and migration through down regulating total EGFR and AKT protein expressions. Erlotinib also inhibited EGF-induced an increase of proliferative and migrative ability in ARPE-19 cells and clearly suppressed EGF-induced EGFR/AKT proteins phosphorylation and decreased expression of N-cadherin, α-SMA, and vimentin proteins. Similarly, EGFR inhibition by EGFR siRNA significantly affected EGF-induced an increase of cell proliferation, viability, and migration, phosphorylation of EGFR/AKT proteins, and up-regulation of N-cadherin, α-SMA, and vimentin proteins.
Erlotinib and EGFR-knockdown suppress EGF-induced cell viability, proliferation, and migration EGFR/AKT pathway in RPE cells. EGFR inhibition may be a possible therapeutic approach for proliferative vitreoretinopathy (PVR).
探讨厄洛替尼和表皮生长因子受体（EGFR）小干扰RNA（siRNA）抑制表皮生长因子受体（EGFR）对表皮生长因子（EGF）诱导的视网膜色素上皮（RPE）细胞激活的影响。
用100 ng/mL EGF激活人RPE细胞系（ARPE-19细胞）。用厄洛替尼和EGFR siRNA干预EGF处理。分别采用甲基噻唑基四氮唑（MTT）法、溴脱氧尿苷（BrdU）染色法和伤口愈合试验检测细胞活力、增殖和迁移情况。通过蛋白质印迹法检测EGFR/蛋白激酶B（AKT）信号通路蛋白以及N-钙黏蛋白、α-平滑肌肌动蛋白（α-SMA）和波形蛋白。也通过免疫荧光染色检测EGFR。
EGF处理24小时可显著提高ARPE-19细胞的活力、增殖和迁移能力，使EGFR/AKT蛋白磷酸化，并降低总EGFR表达。厄洛替尼通过下调总EGFR和AKT蛋白表达来抑制ARPE-19细胞的活力、增殖和迁移。厄洛替尼还抑制EGF诱导的ARPE-19细胞增殖和迁移能力增加，并明显抑制EGF诱导的EGFR/AKT蛋白磷酸化，降低N-钙黏蛋白、α-SMA和波形蛋白的表达。同样，EGFR siRNA抑制EGFR可显著影响EGF诱导的细胞增殖、活力和迁移增加，EGFR/AKT蛋白磷酸化，以及N-钙黏蛋白、α-SMA和波形蛋白的上调。
厄洛替尼和EGFR基因敲低可抑制EGF诱导的RPE细胞活力、增殖和迁移以及EGFR/AKT信号通路。抑制EGFR可能是增殖性玻璃体视网膜病变（PVR）的一种潜在治疗方法。
