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目的:检测合成的蛋白靶向嵌合分子(EGF-PROTAC)通过泛素-蛋白酶体途径体外对人鼻咽癌CNE-2细胞表皮生长因子受体(EGFR)靶向降解作用,并证实对该细胞增殖、凋亡的影响。方法:EGF-PROTAC处理细胞后,应用Western blot技术分析EGFR蛋白表达水平;CKK-8、流式细胞术、Transwell侵袭实验检测EGF-PROTAC对CNE-2产生的生物学效应。结果:Western blot技术显示EGF-PROTAC组EGFR蛋白的表达水平显著低于对照组(P<0.05);CKK-8法显示EGF-PROTAC组3、6、9、12hCNE-2细胞存活率显著低于溶剂对照组(P<0.05);流式细胞术显示EGF-PROTAC组细胞凋亡指数显著高于对照组(P<0.05);Transwell法显示EGF-PROTAC组CNE-2细胞侵袭到Transwell小室滤膜下细胞数显著低于溶剂对照组(P<0.05)。结论:EGF-PROTAC能够靶向降解CNE-2细胞的EGFR,在体外能抑制鼻咽癌细胞的生长、促进凋亡。
OBJECTIVE: To detect the targeted degradation of human epidermal growth factor receptor (EGFR) on human nasopharyngeal carcinoma cell line CNE-2 by the ubiquitin-proteasome pathway in vitro and to demonstrate that EGF- Cell proliferation and apoptosis. METHODS: The EGFR protein expression was analyzed by Western blot after treated with EGF-PROTAC. The biological effects of EGF-PROTAC on CNE-2 production were detected by CKK-8, flow cytometry and Transwell invasion assay. Results: Western blot showed that the expression of EGFR protein in EGF-PROTAC group was significantly lower than that in control group (P <0.05). CKK-8 assay showed that the survival rate of 3, 6, 9, The apoptosis index of EGF-PROTAC group was significantly higher than that of the control group (P <0.05) by flow cytometry. Transwell assay showed that the invasion of CNE-2 cells in EGF-PROTAC group to Transwell cell membrane The number of cells was significantly lower than the solvent control group (P <0.05). Conclusion: EGF-PROTAC can target the degradation of EGFR in CNE-2 cells and inhibit the growth of nasopharyngeal carcinoma cells in vitro and promote apoptosis.