c-MET抑制剂联合EGFR-TKI对耐药肺癌细胞增殖、凋亡的影响及其机制

Effects of c-MET inhibitor combined with EGFR-TKI on proliferation and apoptosis of drug-resistant lung cancer cells

目的观察c-MET抑制剂SU11274联合表皮生长因子-酪氨酸激酶抑制剂(EGFR-TKI)吉非替尼或埃罗替尼对人肺癌细胞株PC9/R(PC9细胞的获得性吉非替尼EGFR-TKI耐药株)增殖、凋亡的影响,并探讨其作用机制。方法培养人肺癌细胞株PC9/R,将PC9/R细胞分为对照组(C组)、SU11274组(S组)、吉非替尼组(G组)、埃罗替尼组(E组)、吉非替尼联合SU11274组(GS组)、埃罗替尼联合SU11274组(ES组)。S组、G组、E组、GS组、ES组分别加入SU11274 5μmol/L、吉非替尼1μmol/L、埃罗替尼1μmol/L、吉非替尼1μmol/L+SU11274 5μmol/L、埃罗替尼1μmol/L+SU11274 5μmol/L,C组不加药物。于给药72 h后采用MTT法检测细胞增殖活力,流式细胞仪技术检测细胞凋亡情况。采用Western blotting法检测各组细胞中的p-EGFR、p-AKT、p-STAT3蛋白。结果GS组、ES组细胞存活率分别低于G组、E组(P均<0.05),联合组细胞存活率均低于S组(P均<0.05),G组、E组、GS组、ES组细胞存活率均低于C组(P均<0.05)。GS组、ES组早期细胞凋亡比例分别高于G组、E组(P均<0.05),联合组早期细胞凋亡比例均高于S组(P均<0.05),GS、ES、S组早期细胞凋亡比例均高于C组(P均<0.05)。GS组、ES组p-EGFR、p-AKT、p-STAT蛋白相对表达量分别低于G组、E组(P均<0.05),联合组p-EGFR、p-AKT、p-STAT蛋白相对表达量均低于S组(P均<0.05),G组、E组、GS组、ES组蛋白相对表达量均低于C组(P均<0.05)。结论 SU11274联合吉非替尼或埃罗替尼可抑制EGFR-TKI耐药的肺癌细胞增殖并促进其凋亡,作用机制可能与抑制EGFR信号通路功能有关。

Objective To observe the effects of c-MET inhibitor SU11274 combined with the epidermal growth factor receptor-tyrosine kinase inhibitor（ EGFR-TKI） gefitinib or erlotinib on the proliferation and apoptosis of human lung cancer cell line PC9/R（ acquired gefitinib EGFR-TKI-resistance cell line PC9 cells） and to investigate the mechanism. Methods We cultured the human lung cancer cell line PC9/R and divided them into six groups： the control group（ group C）,SU11274 group（ group S）,gefitinib group（ group G）,erlotinib group（ group E）,gefitinib combined with SU11274 group（ group GS）,and erlotinib combined with SU11274 group（ group ES）. The groups S,G,E,GS,and ES were added with5 umol/L SU11274,1 umol/L gefitinib,1 umol/L erlotinib,1 μmol/L gefitinib ＋ 5 μmol/L SU11274,and 1 μmol/L erlotinib ＋ 5 μmol/L SU11274; group C was not added any drugs. At 72 h after administration,the cell proliferation was detected by using MTT method,and the apoptosis was detected by using flow cytometry. The protein expression of p-EGFR,p-STAT3,and p-AKT was detected by Western blotting. Results The cell survival rates were lower in the groups GS and ES than in the groups G and E,and the cell survival rates of the combination groups were lower than that of group S（ all P〈0. 05）. The cell survival rates of groups G,E,GS,and ES were lower than that of group C（ all P〈0. 05）,The early apoptosis rates of the groups GS and ES were higher than those of the groups G and E respectively,and both groups were lower than group S（ all P〈0. 05）. The early apoptosis rates of groups GS,ES,and S were higher than that of the group C（ all P〈0. 05）. The protein expression of p-EGFR,p-STAT3,and p-AKT of the groups GS and ES were lower than those of the groups G and E,respectively（ all P〈0. 05）. The protein expression of p-EGFR,p-STAT3,and p-AKT of the combination group was lower than that of the group S,and the protein expression of p-EGFR,p-STAT3,and p-AKT of the groups G,E,GS,and ES was lower than that of the group C（ all P〈0. 05）. Conclusions SU11274 combined with gefitinib or erlotinib can inhibit the proliferation and promote apoptosis of the EGFR-TKI resistance lung cancer cells by inhibiting the function of EGFR pathway.