| 肺腺癌表皮生长因子受体T790M突变裸鼠模型的建立与评价 |
| 投稿时间：2016-03-10 修订日期：2016-03-15 点此下载全文 |
| 引用本文：李亚清,应希旺,严建平,曹立名,邬盛昌,刘元顺.肺腺癌表皮生长因子受体T790M突变裸鼠模型的建立与评价[J].医学研究杂志,2016,45(10):72-75 |
| DOI：
10.11969/j.issn.1673-548X.2016.10.018 |
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| 基金项目:国家自然科学基金资助项目（81470109）；浙江省公益性技术应用研究计划基金资助项目（2014C37022） |
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| 中文摘要:目的 建立表皮生长因子受体（EGFR）T790M突变的肺腺癌裸鼠模型，为探讨肺腺癌EGFR-TKIs 的继发性耐药机制提供工具。方法 建立对吉非替尼耐药的人肺腺癌细胞株RPC-9，以皮下异位移植法建立EGFR T790M突变肺腺癌裸鼠模型。进行组织病理学分析，以基因测序法检测EGFR基因突变；以Western blot法检测EGFR及磷酸化EGFR表达水平。结果 吉非替尼浓度增到10μmol/L时，基因测序法检测显示PC-9细胞EGFR基因E20发生突变，其氨基酸变化为790T>T/M。在4周内，PC-9移植组及RPC移植组肿瘤体积明显差异；移植后第5周时，RPC移植组肿瘤体积为944±20mm3，PC移植组肿瘤体积为878±8mm3，RPC移植组肿瘤体积显著大于PC-9移植组（P<0.05）。PC-9移植及RPC移植组肿瘤组织病理均为低分化腺癌。RPC-9移植组肿瘤组织EGFR基因存在E20发生突变，其氨基酸变化为790T>T/M。PC-9移植组及RPC移植组肿瘤组织EGFR及Tyr992磷酸化的EGFR表达水平差异均无统计学意义（P>0.05）。结论 通过皮下移植对吉非替尼继发性耐药的人肺腺癌细胞RPC-9成功建立EGFR T790M 突变的肺腺癌裸鼠模型，为研究肺腺癌EGFR-TKIs 的继发性耐药提供了有力工具。 |
| 中文关键词:肺腺癌 表皮生长因子受体 继发性耐药 动物模型 |
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| Development of a Nude Mouse Model with Lung Adenocarcinoma Expressing EGFR T790M Mutants Associated with Secondary Resistance to EGFR Tyrosine Kinase Inhibitors |
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| Abstract:Objective To develop a nude mouse model with lung adenocarcinoma expressing EGFR T790M mutants associated with secondary resistance to EGFR kinase inhibitors. Methods The RPC-9 cells expressing EGFR T790M mutantswill tyrosinebe were established under the continuous selective pressure of gefitinib, and then a subcutaneous xenotransplanted tumor model of human ameloblastoma in nude mice will be developed using RPC-9. The histopathological changes were analyzed. EGFR gene mutations were detected by gene sequencing. The expression levels of EGFR and phosphorylated EGFR were detected by Western blot. Results A resistant subline designated RPC-9 from PC-9 cells was established, which was able to grow in the presence of 10μmol/L of gefitinib. There was the mutated EGFR (exon 20) carried 790T>T/M mutation detected using gene sequencing in RPC-9 cells. There were no significant differences in the xenotransplanted tumor sizes between the PC-9 group and the RPC group in 4 weeks (P>0.05). In the fifth week after the xenotransplantation, the xenotransplanted tumors in the RPC group became larger than those in the PC group (P<0.05).They was lung adenocarcinoma in both the PC-9 group and the RPC group. There was the mutated EGFR (exon 20) carried 790T>T/M mutation detected using gene sequencing in the tumor tissues of RPC-9 group. There was no significant difference in expression levels of EGFR and the Tyr992 phosphorylated EGFR between the PC-9 group and the RPC group (P>0.05). Conclusion A subcutaneous xenotransplanted tumor model of human ameloblastoma in nude mice was successfully developed, in which the xenotransplanted tumors expressing EGFR T790M mutants associated with secondary resistance to EGFR tyrosine kinase inhibitors. |
| keywords:Lung adenocarcinoma Epidermal growth factor receptor Secondary drug resistance Animal model |
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