除痰解毒方通过EMT途径逆转肺癌吉非替尼耐药

Chutan-Jiedu decoction reverses resistance of lung cancer to gefitinib by processing EMT

目的:从上皮-间充质转化(EMT)途径探讨除痰解毒方逆转肺癌分子靶向耐药的机制。方法:建立人肺腺癌耐药细胞株H1975裸鼠模型(n=60),随机分为:模型组,吉非替尼(0. 04 g/kg)组,除痰解毒方低、中、高剂量(13. 52 g/kg、27. 04 g/kg和54. 08 g/kg)组,除痰解毒方中剂量(27. 04 g/kg)联合吉非替尼(0. 04 g/kg)组,每组10只。治疗2周,检测瘤体积和瘤质量,计算抑瘤率,采用免疫组织化学法、Western blot法和real-time PCR法检测各组肿瘤组织中E-cadherin、Snail和vimentin蛋白及m RNA的变化。结果:与模型组及吉非替尼组比较,除痰解毒方中、高剂量组及联合用药组瘤体积和瘤质量显著降低(P <0. 05)。免疫组织化学法、Western blot法及realtime PCR法结果均显示除痰解毒方中剂量组和联合用药组E-cadherin蛋白及m RNA的表达水平显著上调,Snail和vimentin蛋白及m RNA表达水平较模型组与吉非替尼组显著下调(P <0. 05)。结论:除痰解毒方可抑制H1975细胞裸鼠移植瘤的生长,逆转肺癌对吉非替尼的耐药,其作用机制可能与调控EMT相关标志蛋白的表达有关。

AIM:To investigate the mechanism of Chutan-Jiedu decoction(CJD)reversing the resistance of lung cancer to gefitinib via epithelial-mesenchymal transition(EMT)pathway.METHODS:BALB/c nude mice(n=60)were selected to establish lung cancer xenograft model with human lung adenocarcinoma drug-resistant cell line H1975,which were randomly divided into 6 groups(10 mice per group):model group,gefitinib(0.04 g/kg)group,low-dose(13.52 g/kg)CJD group,middle-dose(27.04 g/kg)CJD group,high-dose(54.08 g/kg)CJD group,and combined medication group(27.04 g/kg CJD+0.04 g/kg gefitinib).The mice in each group were treated for 2 weeks before the tumor size and tumor weight were detected for the calculation of the tumor inhibitory rate.The mRNA and protein expression levels of E-cadherin,Snail and vimentin were determined by immunohistochemistry,Western blot and real-time PCR.RESULTS:Compared with model group and gefitinib group,the tumor size and the tumor weight in middle-dose CJD group,high-dose CJD group and combined medication group were decreased significantly(P<0.05).The results of immunohistochemistry,Western blot and real-time PCR showed that the expression of E-cadherin at mRNA and protein levels was increased significantly,while the expression of Snail and vimentin at mRNA and protein levels was decreased significantly(P<0.05).CONCLUSION:The growth of lung adenocarcinoma H1975 xenografts in nude mice is inhibited by CJD.In addition,the resistance of lung cancer to gefitinib is reversed.The mechanism may be related to the regulation of EMT-related protein expression.