TGF-β1介导的上皮-间质转分化在Gefitinib耐药中的作用

发布时间：2018-05-18 07:28

本文选题：肺肿瘤 + TGF-β　；参考：《第三军医大学学报》2013年15期

【摘要】：目的调查上皮-间质转化(epithelial-mesenchymal transition,EMT)在非小细胞肺癌(non-small cell lungcancer,NSCLC)患者接受吉非替尼(Gefitinib)治疗反应性中的作用及机制。方法突变富集PCR法检测NSCLC患者EGFR突变状况;免疫组化法检测癌组织中上皮钙粘蛋白(E-cadherin)和纤维连接蛋白(Fibronectin)的表达情况,探讨EMT与表皮生长因子受体酪氨酸激酶抑制剂(epidermal growth factor receptor tyrosine kinase inhibitors,EGFR-TKIs)治疗敏感性的关系。体外选取人肺腺癌细胞系PC9细胞,经TGF-β1反复处理4周,观察细胞形态学变化;MTT检测TGF-β1处理后细胞对Gefitinib敏感性的影响;Western blot验证EMT相关标记蛋白表达变化并检测EGFR信号通路下游蛋白的变化。结果 43例NSCLC标本中,EGFR 19、21外显子突变率为58.14%(25/43)。具有EGFR基因突变的肿瘤E-cadherin的表达水平显著高于EGFR野生型(70.00%vs 30.00%,P0.05)。接受Gefitinib总体有效率为46.51%(20/43),具有E-cadherin阳性表达的患者治疗反应性明显好于Fibronectin阳性的患者(65.00%vs 30.43%,P0.05)。TGF-β1可诱导PC9细胞向间质型细胞形态转化,上调Fibronectin的表达;与亲本PC9细胞相比,TGF-β1处理的细胞对Gefitinib的敏感性下降(P0.05);这种敏感性的下降伴随着AKT和STAT3的持续活化。结论 EMT在EGFR-TKI耐药中发挥着重要作用,TGF-β1诱导的EMT可影响PC9细胞对Gefitinib的敏感性,这种效应可能是通过持续活化AKT和STAT3而发挥作用。
[Abstract]:Objective to investigate the role and mechanism of epithelial-mesenchymal transition (EMT) in the response of patients with non-small cell lung cancer (NSCLC) to gifetinidin (Gefitinib). Methods the mutation of EGFR in NSCLC patients was detected by mutagenic enrichment PCR method, and the expression of E-cadherin and fibronectin in cancer tissues was detected by immunohistochemistry, and the expression of E-cadherin and fibronectin were detected by immunohistochemistry. To investigate the relationship between EMT and the sensitivity of epidermal growth factor receptor tyrosine kinase inhibitors EGFR-TKIs. Human lung adenocarcinoma cell line PC9 was selected in vitro and treated with TGF- 尾 1 for 4 weeks. The effect of TGF- 尾 1 on the sensitivity of cells to Gefitinib was observed. Western blot was used to verify the expression of EMT related marker proteins and the downstream proteins of EGFR signaling pathway were detected. Results in 43 NSCLC specimens, the mutation rate of exon 19 / 21 was 58.14% and 25 / 43% respectively. The expression of E-cadherin in the tumor with EGFR gene mutation was significantly higher than that in the EGFR wild-type tumor (P 0.05). The overall effective rate of Gefitinib was 46.51% 20 / 43. The response of patients with positive E-cadherin expression was significantly better than that of Fibronectin positive patients (65.00 vs 30.43 + P0.05N. TGF- 尾 1). TGF- 尾 1 could induce the morphologic transformation of PC9 cells to interstitial cells and up-regulate the expression of Fibronectin. Compared with parental PC9 cells, the sensitivity of TGF- 尾 _ 1 treated cells to Gefitinib decreased by P0.05G, which was accompanied by the continuous activation of AKT and STAT3. Conclusion EMT plays an important role in EGFR-TKI resistance. TGF- 尾 1-induced EMT may affect the sensitivity of PC9 cells to Gefitinib, which may be due to the continuous activation of AKT and STAT3.
【作者单位】：广州医科大学附属第一医院呼吸科;广州呼吸疾病研究所呼吸疾病国家重点实验室;青岛大学医学院附属医院呼吸科;
【基金】：国家自然科学基金(81200050) 呼吸疾病国家重点实验室青年基金(2011-A8)~~
【分类号】：R329

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