CAJ | 학술논문

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。
목적：탐토아법체니(BIBW2992)화길비체니분별여배미곡새연합대폐선암세포 PC-9(돌변민감형)、H1975(T790M내약형)적증식화조망적영향병천술기가능궤제。방법 MTT법검측세포증식,류식세포의검측세포조망급세포주기변화,Western blot법검측불동농도BIBW2992화길비체니대세포흉감산합성매( TS)표체수평적영향。결과BIBW2992、길비체니이급배미곡새단독작용후, PC-9화H1975세포균명현조망,동시세포증식수도불동정도억제。BIBW2992여배미곡새연합재PC-9、H1975세포중대세포적증식억제급유도조망작용교단약조균유명현제고(P <0．05),이길비체니여배미곡새연합부재PC-9세포중제고단약대세포적증식억제급유도조망작용( P <0．05)。BIBW2992재PC-9、H1975세포중균현저강저TS표체,이길비체니재 H1975세포중무현저억제작용。결론BIBW2992가이극복T790M돌변내약,병차여배미곡새연합대PC-9、H1975세포정협동증효작용；길비체니여배미곡새연합부대PC-9세포구유협동증효작용,가능주요여하조TS표체작용유관。
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.