Molecular Biology Reports, 2012, 39(4):4971-4979.

Synergistic inhibitory effects by the combination of gefitinib and genistein on NSCLC with acquired drug-resistance in vitro and in vivo

Hang Zhu, Hua Cheng, Yuan Ren, et al

Department of Child and Adolescent Health, School of Public Health and Tropical Medicine Southern Medical University Guangzhou, China; et al

In clinical practice, most patients with non small cell lung cancer (NSCLC) who respond to tyrosine kinase inhibitors eventually progress because of an acquired resistance mutation, T790M, in epidermal growth factor receptor (EGFR). Thus, it is important to identify a new drug to reduce resistance. The aim of this study was to test whether genistein combined with gefitinib is effective against NSCLC in a cell line carrying T790M, and to clarify the underlying mechanisms. The human lung cancer cell line H1975 was used as an in vitro and in vivo model. Cells were treated with gefitinib, genistein, or a combination at a range of concentrations. Cell proliferation was calculated to assess the anticancer effects of the compounds in vitro. Flow cytometry and Western blotting were employed to determine the inhibitory effects on proliferation and the induction of apoptosis. The in vivo effects of the compounds were examined using a xenografted nude mouse model for validation. Gefitinib together with genistein enhanced both growth inhibition and apoptosis; however, the greatest synergistic effect was observed at low concentrations. p-EGFR, p-Akt, and p-mTOR expressions in vitro were reduced more by the combined use of the drugs, whereas caspase-3 and PARP activities were increased. Significantly more tumor growth inhibition was detected following combination treatment in the in vivo model. These findings suggest that genistein enhanced the antitumor effects of gefitinib in a NSCLC cell line carrying the T790M mutation. This synergistic activity may be due to increased inhibition of the downstream molecular and pro-apoptotic effects of EGFR.

荷兰《分子生物学报告》，2012年4月

在体内和体外联合使用吉非替尼和金雀异黄素对获得性耐药非小细胞肺癌的协同抑制作用

朱航、 程华、 任远等

中国广州南方医科大学公共卫生学院和热带医学院儿童和青少年健康系等

在临床实践中，大多数患有非小细胞肺癌 (NSCLC)并对酪氨酸激酶抑制剂有反应的患者，由于表皮生长因子受体 (EGFR) 上的T790M发生获得性耐药突变，最终癌症得以发展。因此，确定减少耐药性的新型药物十分重要。本研究的目的是测试金雀异黄素与吉非替尼的联合使用是否能有效地治疗携带 T790M的NSCLC，并阐明其潜在的机制。人体肺癌细胞 H1975 被用作体内和体外的模型。用一系列浓度的吉非替尼、 金雀异黄素或两者的混合物治疗细胞。计算细胞增殖的结果来评估这些化合物在体外的抗肿瘤作用。用流式细胞仪和免疫印迹法来确定其抑制细胞增殖和诱导细胞凋亡的作用。使用裸鼠移植瘤模型考察化合物的体内作用。吉非替尼和金雀异黄素联合增强生长抑制和凋亡作用；而在低浓度时可以观察到最大的协同效应。通过药物的联合使用大幅度降低了p-EGFR，p-Akt和p-mTOR在体外的 表达，而半胱氨酸蛋白酶-3 和 PARP的活性被增强。在体内模型中经过联合治疗后检测到更显著的抑制肿瘤生长的作用。这些结果表明，染料木黄酮增强了吉非替尼对携带 T790M 突变非小细胞肺癌的抗肿瘤作用。这种协同作用可能是因为抑制下游分子和 EGFR 促凋亡作用的增强。