摘要
目的探讨芬维A胺[fenretinide,N-(4-hydroxyphenyl)retinamide(4-HPR),一种人工合成的维甲酸]与硼替佐米联合应用对非小细胞肺癌A549细胞的促凋亡作用。方法将非小细胞肺癌细胞株A549用不同浓度的4-HPR(2.5、5、10、20μmol/L)和硼替佐米(0.1、0.2、0.4、0.8μmol/L)单独或联合处理24h。应用MTT法检测4-HPR和硼替佐米单独或联合处理对细胞的生长抑制作用;碘化丙啶(PI)染色和流式细胞术检测细胞周期;AnnexinⅤ-FITC和PI双染检测细胞凋亡;实时荧光定量PCR和蛋白质印迹检测内质网应激相关凋亡蛋白CHOP的表达。结果 4-HPR和硼替佐米单独处理肺癌细胞株A549能够以剂量依赖性方式抑制细胞增殖,两药联用后抗增殖作用明显增强。细胞周期检测显示两药联用能导致细胞停滞于G0/G1期,S期细胞显著减少。与两药单独使用相比,4-HPR和硼替佐米联用能显著增强A549细胞凋亡,伴随内质网应激蛋白CHOP的mRNA和蛋白表达增强。结论 4-HPR和硼替佐米联用能促进肺癌A549细胞的凋亡,为药物联合治疗肺癌提供了实验基础。
Objective To explore the synergistic apoptosis-promoting effect of fenretinide （N-[4-hydroxyphenyl] retinamide, 4-HPR, a synthetic retinoic acid） with bortezomib in non-small cell lung cancer （NSCLC） A549 cells. Methods NSCLC A549 cells were treated with 4-HPR and bortezomib alone or in combination at different concentrations （2.5, 5,10 and 20 Mmol/L for 4-HPR; 0.1,0.2,0. 4 and 0.8 pmol/L for bortezomib） for 24 h. MTT assay was performed to detect cell growth inhibition. Propidium iodide （PI） staining and flow cytometry were performed to analyze cell cycle. Annexin V- FITC and PI double staining was performed to detect apoptosis. Real-time quantitative PCR and Western blotting analysis were performed to examine the expression of endoplasmic reticulum stress protein CHOP. Results 4-HPR or bortezomib alone inhibited the cell proliferation in a dose-dependent manner, and combined treatment with both 4-HPR and bortezomib showed significantly a stronger anti-proliferative effect. Cell cycle analysis showed that the combination of the two drugs caused cell cycle arrest in the G0/G1 phase, with S phase cells significantly reduced. Compared with 4-HPR or bortezomib used alone, combination of both significantly enhanced the apoptosis of A549 cells, accompanied by enhanced expression of CHOP mRNA and protein, an endoplasmic reticulum stress marker. Conclusion Combination of 4-HPR and bortezomib can promote apoptosis in lung cancer A549 cells, which provides an experimental basis for their combination treatment of lung cancer.
出处
《第二军医大学学报》
CAS
CSCD
北大核心
2013年第11期1177-1182,共6页
Academic Journal of Second Military Medical University
基金
国家自然科学基金(81000006)~~
