联烯化合物PHPO对肺癌细胞株A549增殖及凋亡的影响

Effects of PHPO on apoptosis and proliferation of lung cancer cell line A549

目的·探讨新型联烯化合物(1-苯基-1,2-丙二烯-1-基)二苯基氧膦(PHPO)对肺癌细胞株A549增殖和凋亡的影响。方法·用不同浓度的PHPO处理A549细胞,利用CCK-8法和流式细胞术检测PHPO对细胞增殖、细胞凋亡和细胞周期的影响,利用划痕实验检测PHPO对细胞迁移能力的影响,real-time PCR检测细胞凋亡及细胞周期相关基因的表达,Western blotting检测丝裂原活化蛋白激酶(MAPK)通路中相关蛋白表达及其磷酸化水平的情况。建立肺癌A549裸鼠移植瘤模型,每日注射PHPO进行治疗,观察肿瘤体积的变化。结果·与对照组相比,PHPO能显著降低A549细胞株的增殖活性,其24 h的IC_(50)值为44.23μmol/L;同时,PHPO干预能使A549细胞的凋亡率升高、迁移能力下降;促凋亡蛋白Bax和周期调控蛋白P21 mRNA表达水平升高,抗凋亡蛋白Bcl-2 m RNA表达水平显著降低(均P<0.05);p-p38、p-ERK和p-JNK蛋白表达增加;注射PHPO能抑制移植瘤小鼠肿瘤的生长,与对照组相比有显著差异(P<0.05)。结论·PHPO对A549细胞具有抑制增殖的作用,能诱导细胞凋亡并发生G1期细胞阻滞,降低肺癌细胞的迁移能力,其机制可能与PHPO激活MAPK信号通路,促进p38、ERK和JNK磷酸化有关。

Objective · To investigate the effect of a new type of allene compound, 1-phenylpropadienyl phosphine oxide （PHPO）, on proliferation and apoptosis of lung cancer cell line A549. Methods · A549 cells were treated with different concentrations of PHPO. The effects of PHPO on cell proliferation, apoptosis and cell cycle were detected by CCK-8 and flow cytometry assay. Wound healing test was used to measure the migration ability of A549 cells. Real-time PCR was used to detect the expression of apoptosis and cell cycle related gene. The expression of proteins in MAPK pathway was assayed by the Western blotting. The nude mice xenograft model of human lung cancer A549 cells was established. After tumor formation, PHPO was injected daily for treatment, and the tumor size was observed. Results · Compared to the control group, PHPO significantly inhibited the cell viability of A549 cells and induced apoptosis of them, and the IC50 value of 24 h is 44.23 μmol/L. PHPO blocked the cell cycle in the G1 phase significantly. The migration capacity of PHPO-treated cells was decreased. The mRNA levels of Bax and P21 were up-regulated in PHPO-treated group, and the mRNA lever of Bcl-2 was down-regulated （P〈0.05）. PHPO increased the phosphorylation levels of p38, ERK and JNK. Injection of PHPO could significantly inhibit the growth of tumor in the xenograft model compared to the control group （P〈0.05）. Conclusion · PHPO can induce the apoptosis and inhibit the proliferation of A549 cells, block the cell cycle in the G1 phase and decrease the migration ability of A549 cells significantly. The mechanism may be related to the activation of MAPK signaling pathway by PHPO and the increase of phosphorylation of p38, ERK and JNK.