蟾毒灵逆转肺腺癌H1975细胞对吉非替尼耐药的体内实验研究

Bufalin reverses resistance to ge tinib in xenografts of lung adenocarcinoma H1975 cells in nude mice

目的:观察蟾毒灵联合吉非替尼对肺腺癌H1975细胞裸鼠移植瘤生长的影响,并探讨其可能的作用机制。方法:建立肺腺癌H1975细胞裸鼠皮下移植瘤模型(共40只)后,随机分为模型组(不进行药物干预)、吉非替尼组、蟾毒灵组及吉非替尼和蟾毒灵联合用药组(每组10只),分别用相应药物干预3周,计算肿瘤抑制率;应用TUNEL法检测移植瘤细胞凋亡,蛋白质印迹法检测移植瘤组织表皮生长因子受体-磷酯酰肌醇-3激酶(epidermal growth factor receptor-phosphoinositide 3-kinase,EGFR-PI3K)/Akt信号通路相关蛋白的表达。结果:实验结束时,模型组、吉非替尼组、蟾毒灵组和联合用药组的抑瘤率分别为0%、16.14%、33.48%和60.39%。模型组、吉非替尼组、蟾毒灵组和联合用药组的移植瘤组织中肿瘤细胞凋亡率分别为(13.11±1.60)%、(15.48±0.43)%、(45.09±3.81)%和(75.8±3.16)%,联合用药组与模型组、吉非替尼组和蟾毒灵组比较,差异有统计学意义(P<0.01)。联合用药组移植瘤组织中p-EGFR、p-PI3K和p-Akt蛋白的表达水平明显下调(P<0.01)。结论:蟾毒灵联合吉非替尼可逆转肺腺癌H1975裸鼠移植瘤对吉非替尼的耐药,其作用机制可能与阻断EGFR-PI3K/Akt信号通路有关。

Objective： To investigate the effect of bufalin in combination with gefitinib on the growth of xenotransplants of lung adenocarcinoma H1975 cells in nude mice, and to explore its possible mechanism. Methods： Forty nude mice bearing subcutaneous xenotransplants of lung adenocarcinoma H1975 cells were randomly divided into four groups （in each group there were 10 nude mice）： control group （no treatment）, gefitinib-treated group, bufalin-treated group and bufalin in combination with gefitinib-treated group. Each group received corresponding intervention for 3 weeks. Tumor inhibitory rate was calculated. Apoptotic rate was measured by TUNEL staining method. The expression levels of EGFR-PI3K （epidermal growth factor receptor-phosphoinositide 3-kinase）/Akt signaling pathway-related proteins in xenografts were detected by Western blotting. Results： The tumor inhibitory rates of the control, gefitinib-treated, bufalin-treated and bufalin in combination with gefitinib-treated groups were 0%, 16.14%, 33.48% and 60.39%, respectively. The apoptotic rate of bufalin in combination with gefitinib- treated group was （75.8±3.16）%, which was significantly higher than those of the control group [（13.11±1.60）%], gefitinib-treated group [（15.48±0.43）%] and bufalin-treated group [（45.09±3.81）%] （P 〈 0.01）. The expression levels of phospho-EGFR, phospho-PI3K and phospho-Akt proteins were significantly reduced in group of bufalin in combination with gefitinib （P 〈 0.01）. Conclusion： Bufalin in combination with gefitinib may block the EGFR-PI3K/Akt pathway to reverse the resistance to gefitinib in xenografts of lung adenocarcinoma H1975 cells in nude mice.