抗体封闭细胞表面GRP78抑制厄洛替尼耐药性口腔舌鳞状细胞癌细胞的生长和转移

Blockade of Cell Surface GRP78 Inhibits the Growth and Metastasis of Erlotinib-Resistant Tongue Squamous Cell Carcinoma Cells

目的观察应用特异性抗体封闭细胞表面葡萄糖调节蛋白78(GRP78)对厄洛替尼耐药的口腔舌鳞状细胞癌(简称鳞癌)细胞生长的抑制作用。方法应用GRP78特异性抗体处理厄洛替尼耐药性舌鳞癌细胞(TCA-8113-ER),采用MTT方法和克隆形成实验检测抗体封闭对TCA-8113-ER细胞增殖情况及其对厄洛替尼反应性的影响,in-cell Western技术检测GRP78在TCA-8113-ER细胞表面的表达,流式细胞术检测TCA-8113-ER细胞凋亡情况,划痕法和Transwell法检测TCA-8113-ER细胞侵袭和转移情况,Western blotting检测c-MET、AKT和STAT3表达及磷酸化水平。结果GRP78特异性抗体可以抑制TCA-8113-ER细胞的生长,促进其凋亡,增强其对厄洛替尼的反应性,抑制其侵袭和转移,抑制c-MET、AKT和STAT3的磷酸化。结论抗体封闭细胞表面GRP78能够抑制厄洛替尼耐药性舌鳞癌细胞的生长、侵袭和转移,增加其对厄洛替尼的敏感性。

Objective To explore the inhibitory effect of a blocking antibody specific to cell surface glucose-regulated protein 78(GRP78)on growth and metastasis of erlotinib-resistant tongue squamous cell carcinoma cells.Methods GRP78-specific antibody was used to treat an erlotinib-resistant cell line established from a tongue squamous cell carcinoma(TCA-8113-ER).MTT and clone formation assays were used to assess the effect of the antibody blocking on the proliferation of TCA-8113-ER cells and their response to erlotinib.Cell surface expression of GRP78 in these cells was analyzed by in-cell Western assay.Apoptosis of TCA-8113-ER cells was evaluated by flow cytometry.Invasive and metastatic potential of TCA-8113-ER cells was analyzed by scratch wound healing and transwell migration assays.Expression levels and phosphorylation states of c-MET,AKT,and STAT3 were analyzed by Western blotting.Results GRP78-specific antibody inhibited the growth and migration of TCA-8113-ER cells while inducing their apoptosis and enhancing their sensitivity to erlotinib.At the molecular level,the treatment inhibited phosphorylation of c-MET,AKT,and STAT3.Conclusion Antibody-mediated blockade of cell surface GRP78 inhibits the growth,invasiveness,and metastatic potential of erlotinib-resistant tongue squamous cell carcinoma cells while increasing their sensitivity to erlotinib.