论文部分内容阅读
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 investigate the pro-apoptotic effects of fenretinide (N- (4-hydroxyphenyl) retinamide (4-HPP), a synthetic retinoid) and bortezomib on A549 cell proliferation in non-small cell lung cancer. Methods Human non-small cell lung cancer cell line A549 was treated with different concentrations of 4-HPR (2.5, 5, 10 and 20 μmol / L) and bortezomib (0.1, 0.2, 0.4 and 0.8 μmol / L) The inhibition of cell growth by 4-HPR and bortezomib alone or in combination was detected by MTT assay. The cell cycle was detected by propidium iodide (PI) staining and flow cytometry. Apoptosis was detected by AnnexinⅤ-FITC and PI double staining Real-time fluorescence quantitative PCR and Western blotting were used to detect the expression of endoplasmic reticulum stress-related apoptosis protein CHOP. Results 4-HPR and bortezomib alone could inhibit the proliferation of lung cancer cell line A549 in a dose-dependent manner. The anti-proliferative effects of the two drugs were significantly enhanced. Cell cycle test showed that the combination of the two drugs can lead to cell arrest in the G0 / G1 phase, S phase cells significantly reduced. Compared with the two drugs alone, 4-HPR combined with bortezomib significantly increased the apoptosis of A549 cells, accompanied by increased expression of the endoplasmic reticulum stress protein CHOP mRNA and protein. Conclusion The combination of 4-HPR and bortezomib can promote the apoptosis of lung cancer A549 cells and provide the experimental basis for the combination therapy of drugs and lung cancer.