毒胡萝卜素联合吉非替尼改善人肺腺癌细胞PC9/GR的耐药性研究

Improvement of gefitinib-resistance of PC9/GR by thapsigargin combined with gefitinib

目的研究毒胡萝卜素(thapsigargin)联合吉非替尼(gefitinib)对人肺腺癌耐药细胞株PC9/GR增殖的影响并探讨可能的机制。方法吉非替尼单独用药或吉非替尼与毒胡萝卜素联合应用,通过CCK8实验检测上述两组药物对PC9/GR细胞增殖的影响;应用流式细胞术鉴定两组药物对PC9/GR细胞凋亡的影响;应用Western blotting法检测两组药物对PC9/GR细胞蛋白ATF-6和IRE1α表达的影响。结果细胞增殖实验显示,与吉非替尼单独用药相比,联合用药组中PC9/GR的增殖受到更强的抑制作用;凋亡实验显示,相较于吉非替尼单独用药,联合用药能够进一步促进细胞的凋亡;Western blotting法显示,与吉非替尼单独用药相比,联合用药后PC9/GR中ATF-6和IRE1α蛋白(内质网应激标志物)表达上调,其差异具有统计学意义。结论在毒胡萝卜素诱导下,PC9/GR细胞对吉非替尼的敏感性增加,其中机制可能与内质网应激有关。

Objective To study the effect and mechanism of the thapsigargin combined with gefitinib on the proliferation of human lung adenocarcinoma gefitinib resistance cell line PC9/GR.Methods The cell viability of PC9/GR treated with gefitinib alone or gefitinib combined with thapsigargin was evaluated by CCK8 assay.The flow cytometry was used to analyze the PC9/GR cell apoptosis indued by the two group drugs.The ATF-6 and IRE1αprotein expression of PC9/GR cells treated with the two group drugs were detected by Western blotting.Results The group of drug combination exhibited enhanced ability to inhibit cell proliferation,promote cell apoptosis and upregulate the ATF-6 and IRE1αprotein expression of the PC9/GR compared with the group gefitinib used alone.Conclusion The sensitivity of PC9/GR to gefitinib was increased when the cells were treated by thapsigargin,which may be related with the state of endoplasmic reticulum stress(ERS)induced by thapsigargin.