盐酸埃克替尼对肺癌细胞增殖和凋亡的影响

Effects of icotinib hydrochloride on the proliferation and apoptosis of human lung cancer cell lines

目的探讨盐酸埃克替尼对不同肺癌细胞系细胞增殖和凋亡的影响。方法运用0～40μmol／L浓度的盐酸埃克替尼和吉非替尼分别处理体外培养的人肺癌细胞系HCC827、H1650、H1975、A549及人表皮癌细胞系A431，应用四甲基偶氮唑蓝比色法观察细胞增殖活力的改变，用流式细胞仪检测药物对细胞凋亡的影响，同时应用免疫印迹法观察下游信号通路的改变。结果盐酸埃克替尼作用于A431和HCC827细胞系的半数抑制浓度分别为（0．04±0．02）、（0．15±0．06）txmol／L。盐酸埃克替尼与吉非替尼相比，对A431、HCC827、H1650、H1975和A549细胞系的抑制作用差异均无统计学意义（均P〉0．05）。与H1650、H1975和A549细胞系相比，盐酸埃克替尼可显著抑制A431和HCC827细胞系的生长（P值分别为0．009、0．005、0．000和0．001、0．001、0．000）。盐酸埃克替尼可以通过抑制人表皮生长因子受体（EGFR）磷酸化活性使磷酸化的蛋白激酶B（AKT）、胞外信号调节激酶（ERK）和凋亡抑制基因survivin蛋白表达下调。结论盐酸埃克替尼可通过抑制AKT．ERK通路以及survivin通路的激活，特异性控制EGFR突变或EGFR过表达的肺癌细胞系的增殖并促进其凋亡。

Objective To explore the effects of icotinib on the proliferation and apoptosis of various lung cancer cell lines. Methods Human lung cancer cell lines HCC827, H1650, H1975, A549 and human epidermal cancer cell line A431 were treated in vitro with icotinib or gefitinib at a concentration gradient of 0 -40 μmol/L. Their proliferation effects were analyzed by the thiazoiyl blue （MTY） assay and the apoptotic effects detected by flow cytometer. The downstream signaling proteins were detected by Western blot. Results The median inhibitory concentrations （ICso） of icotinib for A431 and HCC827 cell lines were （0. 04 ± 0. 02） and （0. 15 ± 0. 06） μmol/L respectively. No significant differences existed between the inhibitions of gefitinib and icotinib on A431, HCC827, H1650, H1975 and A549 cell lines （ all P 〉 0. 05）. Compared with H1650, H1975 and A549 cell lines, icotinib significantly inhibited A431 （ P = 0. 009, 0. 005 and 0. 000） and HCC827 （P -0. 001, 0. 001 and 0. 000） cell lines. And it lowered the expressions of p-AKT, p-ERK and survivin protein expression through the inhibited activity of p-EGFR protein. Conclusion Icotinib can arrest the proliferation of lung adenocarcinoma cells with EGFR mutation or overexpression by inhibiting the signal pathways of AKT-ERK and survivin.