【作者】 袁渊；

【导师】 束永前；

【作者基本信息】 南京医科大学 ， 肿瘤学， 2008， 硕士

【摘要】 目的:探讨雌激素受体拮抗剂TAM联合表皮生长因子受体抑制剂gefitinib对人NSCLC细胞株增殖和凋亡的影响及相关的分子机制,为联合分子靶向和激素治疗NSCLC提供理论依据。方法:以gefitinib及TAM单独或联合处理NSCLC细胞株,MTT法分析细胞增殖,流式细胞仪检测细胞凋亡。应用荧光定量PCR及Western Blot检测三种肺癌细胞株中EGFR及ERβ的表达情况,并分别检测17β-E₂,TAM对细胞中EGFR,以及EGF和gefitinib对ERβ表达的影响,同时还检测了17β-E₂对EGFR的磷酸化作用。结果:联合gefitinib和TAM增强对EGFR,ERβ阳性的A549,H1650细胞的抑制作用,促进凋亡,但对EGFR,ERβ阴性的H520细胞无明显改变。17β-E₂可促进细胞EGFR磷酸化,但抑制其表达,而TAM上调EGFR的表达。同时,EGF及gefitinib对ERβ的表达也有类似的相反的调控作用。结论:联合gefitinib和TAM能够增强对EGFR,ERβ双阳性的肺癌细胞的抑制增殖作用,这可能与细胞内EGFR信号转导途径和雌激素信号通路的交叉有关。因此,EGFR的分子靶向治疗联合激素治疗可能是肺癌治疗的一种有效途径。更多还原

【Abstract】 Objective: To identify the effect of oestrogen receptor (ER) antagonist, tamoxifen, combination with EGFR tyrosine kinase inhibitor, gefitinib, on the proliferation and apoptosis in non-small cell lung cancer (NSCLC) cell lines and the related molecular mechanism, providing theoretical evidence for the molecular targeted treatment combination with endocrine treatment in NSCLC. Methods: The cell lines were treated with gefitinib and tamoxifen alone or in combination. We detected the cell proliferation with MTT, the cell apoptosis with flow cytometry and the expression of EGFR and ERβwith real-time PCR and western-bolt. In addition, we detect the effect of oestrogen and TAM on the expression of EGFR, the effect of EGF and gefitinib on the expression of ERβ, respectively. The effect of oestrogen on the phospho-EGFR was also detected. Results: The combination of tamoxifen and gefitinib increased the inhibition and apoptosis in EGFR and ERβpositive cell lines A549 and H1650, while showed no significant changes in EGFR and ER negative cell line H520. It was found that oestrogen can promote the phospho-EGFR protein expression, meanwhile, EGFR protein expression was down-regulated in response to estrogen and up-regulated in response to Tamoxifen in vitro. Conversely, ERβexpression was decreased in response to EGF and increased in response to gefitinib. Conclusion: The combination of tamoxifen and gefitinib can promote its antiproliferative effects on EGFR and ERβpositive lung cell lines. These results suggested that this might be involved in cross-signaling between the EGFR/ER pathways in NSCLC. In conclusion, it provided rational to combine gefitinib with hormone therapy for lung cancer treatment.更多还原