阿法替尼和吉非替尼分别联合培美曲塞对人肺腺癌细胞作用的研究  

The inhibitory effects of the combination of pemetrexed and BIBW2992 or gefitinib respectively on human lung adenocarcinoma cell line

作　　者：边劲[1] 王琳[1] 寻琛[1] 黄伟[1] 

机构地区：[1]安徽医科大学解放军八一临床学院(中国人民解放军八一医院)肿瘤内科,南京210002

出　　处：《安徽医科大学学报》2014年第6期772-776,共5页Acta Universitatis Medicinalis Anhui

基　　金：南京军区医学科技创新课题项目(编号:ZD19)

摘　　要：目的探讨阿法替尼(BIBW2992)和吉非替尼分别与培美曲塞联合对肺腺癌细胞PC-9(突变敏感型)、H1975(T790M耐药型)的增殖和凋亡的影响并阐述其可能机制。方法 MTT法检测细胞增殖,流式细胞仪检测细胞凋亡及细胞周期变化,Western blot法检测不同浓度BIBW2992和吉非替尼对细胞胸苷酸合成酶(TS)表达水平的影响。结果 BIBW2992、吉非替尼以及培美曲塞单独作用后,PC-9和H1975细胞均明显凋亡,同时细胞增殖受到不同程度抑制。BIBW2992与培美曲塞联合在PC-9、H1975细胞中对细胞的增殖抑制及诱导凋亡作用较单药组均有明显提高(P<0.05),而吉非替尼与培美曲塞联合仅在PC-9细胞中提高单药对细胞的增殖抑制及诱导凋亡作用(P<0.05)。BIBW2992在PC-9、H1975细胞中均显著降低TS表达,而吉非替尼在H1975细胞中无显著抑制作用。结论 BIBW2992可以克服T790M突变耐药,并且与培美曲塞联合对PC-9、H1975细胞呈协同增效作用;吉非替尼与培美曲塞联合仅对PC-9细胞具有协同增效作用,可能主要与下调TS表达作用有关。Objective To investigate the potential anti-proliferative and pro-apoptotic effects of the combination of pemetrexed and afatinib（BIBW2992） or gefitinib respectively on gefitinib-sensitive cells （PC-9） and gefitinib-re-sistant cells with the T790M mutation of EGFR（H1975） and explore its possible anti-cancer mechanism. Methods MTT assay was performed to reveal the inhibitory effect on cell proliferation. Flow cytometry using annexin V-FITC/PI staining was employed to measure the cell apoptosis and cell cycle. Western blot was performed to detect the protein expressions of thymidylate synthase （ TS） after treated with different dose of BIBW2992 and gefitinib re-spectively . Results The PC-9 and H1975 cells were inhibited to different degrees after treatment with BIBW2992 , gefitinib and pemetrexed alone and had apparent apoptotic features. The combined treatment with BIBW2992 and pemetrexed resulted in a synergistic effect in inducing apoptosis and inhibiting cell proliferation compared with BIBW2992 and pemetrexed alone both on the PC-9 and H1975 cells（P〈0. 05）. The combined treatment with ge-fitinib and pemetrexed resulted in a synergistic effect only on the PC-9 cells（P〈0. 05）, whereas had no such syn-ergistic effect on the H1975 cells. The BIBW2992 decreased the expressions of TS apparently both on the PC-9 cells and H1975 cells, whereas gefitinib had no such effect on the H1975 cells. Conclusion The BIBW2992 can overcome the drug resistance of cells with the T790M mutation of EGFR. The combination of BIBW2992 and peme-trexed has an enhanced antitumor effect on both PC-9 cells and H1975 cells, whereas the combination of gefitinib and pemetrexed has an enhanced antitumor effect only on PC-9 cells, with the possible mechanism of ability to sup-press the expression of TS.

关 键 词：BIBW2992 吉非替尼 肺癌 细胞周期 细胞凋亡 

分 类 号：R734.2[医药卫生—肿瘤] R329.25[医药卫生—临床医学]