目的： 肝细胞生长因子(hepatocyte growth factor, HGF)诱导敏感非小细胞肺癌(non-small cell lung cancer,NSCLC)细胞对表皮生长因子受体酪氨酸激酶抑制剂（epidermal growth facter receptor tyrosine kinase inhibitor, EGFR-TKI）厄洛替尼耐药，本研究旨在探讨c-Met抑制剂SU11274逆转HGF诱导不同EGFR基因型非小细胞肺癌细胞株对厄洛替尼耐药及逆转耐药机制。方法： 选择人NSCLC细胞株PC9（EGFR突变型，敏感株）、H292（EGFR野生型，敏感株）和A549（EGFR野生型，原发性耐药株），应用厄洛替尼和SU11274（1 μmol /L）单独或联合作用于HGF（40 ng/ml）诱导的细胞株，实验分为6组：C组（不加药对照组）、H组（HGF处理）、E组（厄洛替尼处理组）、S组（SU11274处理组）、HE组（HGF+厄洛替尼处理组）、HES组（HGF+厄洛替尼+ SU11274处理组）。MTT法、流式细胞术分别检测不同药物处理对各细胞增殖、凋亡的影响；应用Western blotting检测不同药物处理对各细胞中c-Met及其下游通道Stat3、Akt、Erk1/2蛋白表达水平。结果：厄洛替尼对3种细胞的增殖抑制作用均呈浓度依赖性，HGF处理能够缓解厄洛替尼的增殖抑制作用（P<0.05）；不同浓度厄洛替尼联合SU11274作用于HGF诱导细胞时3种细胞株存活率比厄洛替尼单独作用于HGF诱导细胞时明显降低（P<0.05）；HGS组的细胞凋亡比HG组明显增加（P<0.05）；HGS组的c-Met、Stat3、Akt、Erk1/2活化蛋白量比HG组明显减少。结论： c-Met抑制剂SU11274和厄洛替尼联合应用可逆转HGF诱导不同EGFR基因型非小细胞肺癌细胞对厄洛替尼耐药，其机制可能与抑制HGF活化的c-Met及其下游通道蛋白表达有关。

Objective:To investigate whether c-Met inhibitor SU11274 can reverse resistance to Erlotinib, an epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI), induced by HGF in non-small cell lung cancer cells with different EGFR gene types. Methods: NSCLC cells with different EGFR gene types, including PC9 (EGFR-activating mutant), H292 (EGFR-wild type), and A549 (EGFR-wild type) were utilized in the study. The experiments for each cell line consisted of six different treatment groups: C group (control), H group (HGF), E group (Erlotinib), S group (SU11274), EH group (Erlotinib+HGF), and ESH group (Erlotinib+SU11274+HGF). Their effects on cell survival and apoptosis were measured by MTT assay and flow cytometry (FCM). The activation of c-Met, Stat3, Akt, and Erk1/2 protein were examined by immunoblotting. Results: Erlotinib inhibited growth of the three cells lines in a dose-dependent manner, and the inhibition was effectively blocked by HGF. The presence of SU11274 significantly decreased the survival rates of cells exposed to HGF and Erlotinib (P<0.05). Apoptosis in cells treated with Erlotinib, SU11274, and HGF was also markedly increased compared with these treated with Erlotinib and HGF only (P<0.05). Similarly, the levels of p-Met, p-Stat3, p-Akt, and p-Erk1/2 in the HES group were significantly lower than that in the HE group (P<0.05). Conclusion: SU11274 reversed HGF-induced resistance to Erlotinib in non-small lung cancer cells with different EGFR gene type, likely due to the inhibition of HGF-induced activation of c-Met and its down streams signaling pathways.

国家自然科学基金资助项目(No. 81160291)