异鼠李素抑制耐吉非替尼人肺癌细胞PC9增殖的研究

Isorhamnetin Suppresses the Growth of Gefitinib Resistant Human Lung Cancer PC9 Cells

目的检测异鼠李素对耐吉非替尼的人肺癌细胞PC9增殖的影响。方法通过持续在人肺癌细胞系PC9的培养条件中加入吉非替尼培养直到PC9细胞出现耐药情况,即得到耐药细胞株(PC9-IR)。再将PC9-IR用不同浓度的异鼠李素处理,分别在24,48和72 h后用噻唑蓝(MTT)法检测细胞的生长情况,以正常人外周血单个核细胞(PBMNCs)作为对照组。同时采用蛋白质印迹法检测药物作用之后信号通路的变化,再用克隆分析检测isorhamentin对细胞克隆形成的影响。结果 MTT证实异鼠李素对PC9-IR的生长有明显的抑制作用,24,48,72 h的半数抑制浓度(IC50)分别为212,98,64μmol.L-1,而对PBMNCs72 h的IC50为204μmol.L-1,蛋白质印迹法证实isorhamnetin可以抑制Akt473位的磷酸化,克隆分析证实40μmol.L-1异鼠李素可明显抑制PC9-IR的克隆形成。结论异鼠李素对耐药的人肺癌细胞系(PC9-IR)的生长有抑制作用,可能因为抑制Akt473位的磷酸化水平。

Objective To investigate the role of isorhamnetin on the growth of PC9-IR cells. Methods The gefitinib-resistant PC9（PC9-1R） cells were generated by consistently culturing the PC9 cells with 10% FBS/DMEM contained 0.5 μmol· L^-1 gefitinib. Then, the PC9-IR cells were treated with isorhamnetin at different concentrations. The proliferation was detected by MTI＂ assay after 24,48 and 72 h, respectively, with PBMNCs as control. Western blot assay was used to detect the change of growth signal pathway. Additionally, clonal assay was performed to analyze the clone formation ability of PC9-IR with or without isorhamnetin. Results Isorhamnetin inhibitted the growth of PC9-IR. The ICso at the three time points were 212 μmol · L^-1 （24 h） ,98μmol· L^-1 1（48 h） ,64μmol· L^-1 （72 h） ,and PBMNCs was 204 μmol· L^-1 （72 h）. It was showed that the phosph-Akt was suppressed and the clonal formation ability of PC9-IR was lowered by isorhamnetin. Conclusion Isorhmantin could inhibit the growth of PC9-IR cells through suppressing the phosphorylation of Akt.