葡萄糖调节蛋白78对L858R突变的非小细胞肺癌erlotinib敏感性的影响

Effects of Glucose-regulated Protein 78 on the Sensitivity of Non-small Cell Lung Cancer Cells Harboring the L858R Mutation to Erlotinib

目的观察葡萄糖调节蛋白78(GRP78)对L858R突变非小细胞肺癌对erlotinib敏感性的影响。方法应用基因转染技术干预H3255细胞中GRP78及其突变体的表达,应用MTT方法检测erlotinib对细胞增殖的抑制情况,应用流式细胞术和FITCTUNEL分析凋亡情况,应用免疫印迹技术检测表皮生长因子受体(EGFR)、细胞外信号调节激酶(ERK)的表达及磷酸化水平。结果过表达GRP78及其多肽结合结构域删除的突变体降低H3255细胞对erlotinib的敏感性,抑制erlotinib诱导的细胞凋亡,促进EGFR、ERK的磷酸化。结论 GRP78通过其ATPase结构域促进非小细胞肺癌细胞对erlotinib耐药。

Objective To investigate the effects of glucose-regulated protein 78（GRP78） on erlotinib sensitivity of L858 R mutation-positive non-small cell lung cancer. Methods The expression of GRP78 and its mutants in H3255 cells was altered using a gene transfection technique. The inhibition of cell proliferation by erlotinib was detected by MTT assay. Apoptosis was analyzed by flow cytometry and fluorescein isothiocyanate-terminal deoxynucleotidyl transferase d UTP nick-end labeling（FITC-TUNEL）,and an immunoblotting technique was used to detect the expression and phosphorylation of epidermal growth factor receptor（EGFR） and extracellular signal-regulated kinase（ERK）. Results Overexpression of GRP78 and its deleted peptide-binding domain reduced the sensitivity of H3255 cells to erlotinib,inhibited erlotinib-induced apoptosis,and promoted the phosphorylation of EGFR and ERK. Conclusion GRP78 promotes resistance to erlotinib in non-small cell lung cancer cells through its ATPase domain.