染料木黄酮联合吉非替尼对人非小细胞肺癌细胞株H1975生长的抑制作用研究

THE INHIBITORY EFFECT OF THE COMBINATION OF GENISTEIN AND GEFITINIB ON NON SMALL CELL LUNG CANCER CELL H1975 AND ITS MECHANISM

目的探讨染料木黄酮（Genistein，Gen）与吉非替尼（Gefitinib，Gef）联合应用对人非小细胞肺癌（NSCLC）细胞株H1975增殖与凋亡的影响及其机制。方法采用MTT比色法观察Gen与Gef单用及联用对人非小细胞肺癌株H1975细胞增殖的影响；利用中效方程计算联合指数（CI值）；流式细胞仪检测细胞凋亡率；Westernblotting检测P—EGFR、P—Akt、p-mTOR和p-Erkl／2的蛋白磷酸化水平。结果Gen与Gel两者联合对H1975细胞的抗增殖效应优于Gef组，协同增效作用明显，抑制率升高，并呈现剂量依赖的抗增殖效应，差异显著，具有统计学意义（P〈0．05）；联用不同浓度的Gen和Gef后，联合给予的指数（CI）值随着浓度的增高（G1～G6），抑制效果逐渐增强，cI值均小于1，表现出协同效应；Gen和Gef联用可显著提高细胞凋亡率；与Gef组相比，联用组P—EGFR、p-Akt和p-mTOR磷酸化水平降低；而对P—Erkl／2磷酸化水平却无抑制作用。结论Gen可以增强Gef对H1975细胞的增殖抑制作用并促进肿瘤细胞的凋亡，同时降低p-EGFR、p-AKT、p-mTOR的磷酸化水平，因而可能对非小细胞肺癌细胞的侵袭、转移潜能具有抑制作用。

Objective To investigate the inhibitory effect of the combination of genistein （Gen） and gefitinib （Get＇） on non small cell lung cancer H1975 cell line and its mechanism. Methods The effect of Gen or/and Gef on NSCLC H1975 cells was studied by MTT growth assay. The median-effect equation was used to calculate the combination index （CI）. Apoptosis rate was detected by flow cytometry. The protein phospborylation levels of EGFR, Akt, mTOR and Erkl/2 were assayed by Western blotting. Results Gen combined with Gel showed obviously antiproliferative effects on H1975 cells than Gef alone. The inhibition rate was increased significantly and dose-dependently （P〈0.05）. The results of CI values showed that the more drug concentration increased （G1-G6）, the more inhibitory effect gradually increased. CI values were less than l, indicating that combination of Gen and Gef had synergistic inhibition effect. Gen combined with Gef could significantly enhance apoptosis rate. Compared with Gel group, phosphorylation levels of EGFR, Akt and roTOR were significantly lowered in combination group, but phosphorylation level of Erkl/2 was not inhibited. Conclusion Gen in combination with Gef can enhance the antiproliferation effect and promote apoptosis in H1975 cells. The phosphorylation levels of EGFR, Akt and m TOR were reduced, which may inhibite the invasion and metastasis of cancer cells.