厄洛替尼联合氯喹对EGFR野生型非小细胞肺癌增殖、凋亡及自噬相关蛋白表达的影响  

Effects of erlotinib combined with choroquine on proliferation,apoptosis,and autophagyassociated protein expression of non-small-cell lung cancer with wild-type EGFR

作　　者：宁凯笛 周恬 项永晶[1] NING Kaidi;ZHOU Tian;XIANG Yongjing(The Second Affiliated Hospital of Guiyang College of Traditional Chinese Medicine,Guiyang 550003,China)

机构地区：[1]贵阳中医学院第二附属医院,贵阳550003

出　　处：《山东医药》2018年第34期13-16,共4页Shandong Medical Journal

基　　金：贵州省卫生计生委科学技术基金项目(gzwjkj2015-1-29)

摘　　要：目的观察厄洛替尼联合氯喹对表皮生长因子受体(EGFR)野生型非小细胞肺癌(NSCLC)细胞增殖、凋亡及自噬相关蛋白表达的影响。方法体外培养K-RAS野生型EGFR野生型NSCLC细胞Spc-A1和K-RAS突变型EGFR野生型NSCLC细胞A549,将其分别分为4组,A、B、C组分别加入厄洛替尼、氯喹、厄洛替尼+氯喹各0.1 m L,D组加入等量培养基。培养72 h,采用MTT法观察细胞增殖情况,并计算厄洛替尼、氯喹的联合指数(CI);培养48 h,采用Hoechst33258染色法观察细胞凋亡情况,Western blotting法检测细胞中的自噬微管相关蛋白轻链3(LC3)。结果 A、B、C组细胞存活率均较D组降低(P均<0.05),C组细胞存活率较A、B组降低(P均<0.05),厄洛替尼、氯喹两药联用的CI<1。Spc-A1细胞A、B、C组细胞凋亡率分别为21.1%、12.1%、48.9%,高于D组的8.3%(P均<0.05),A549细胞A、B、C组细胞凋亡率分别为19.3%、13.4%、51.5%,高于D组的10.2%(P均<0.05),两种细胞的C组细胞凋亡率均高于A、B组(P均<0.05);A、B、C组细胞自噬相关蛋白LC3-Ⅰ表达减少而LC3-Ⅱ表达增多(P均<0.05),C组较A、B组细胞自噬相关蛋白LC3-Ⅰ表达减少而LC3-Ⅱ表达增多(P均<0.05)。结论氯喹可增敏厄洛替尼对EGFR野生型NSCLC细胞增殖的抑制作用,且增敏不受K-RAS基因表型限制;其机制可能与氯喹阻断细胞自噬和促进细胞凋亡有关。Objective To observe the effects of erlotinib combined with choroquine on proliferation, apoptosis, and autophagy-associated protein expression of non-small-cell lung cancer （NSCLC） with wild-type epidermal growth factor receptor （EGFR）.Methods Two NSCLC cell lines Spc-A1（wild-type K-RAS wild-type EGFR） and A549 （K-RAS mutant wild-type EGFR） cultured in vitro were divided into four groups. The cells in the groups A, B, and C were added with erlotinib, choroquine, and erlotinib＋choroquine （0.1 mL）, respectively, and cells in the group D were added with the same amount of medium. MTT assay was performed to detect the cell proliferation after 72 hours of culture and the combination index of erlotinib and choroquine was calculated. Hoechst33258 staining was performed to observe apoptosis after 48 hours of culture. The expression of autophagy microtubule-associated protein light chain 3（LC3） was detected by Western blotting.Results The cell survival rates of groups A, B, C were lower than that of the group D （all P 〈0.05）, and that of group C was lower than those of groups A and B （all P 〈0.05）, the combination index of erlotinib and chloroquine was 〈1. The apoptosis rates of Spc-A1 cells in the groups A, B, and C were 21.1%, 12.1%, and 48.9%, respectively, which were higher than that （8.3%） of group D （all P 〈0.05）, the apoptosis rates of A549 cells in the groups A, B, and C were 19.3%, 13.4%, and 51.5%, which were higher than that （10.2%） of group D （all P 〈0.05）, the apoptosis rate of group C was higher than those of groups A and B in both Spc-A1 and A549 cells （all P 〈0.05）, meanwhile, the expression of autophagy-related protein LC3-Ⅰin the groups A, B, and C decreased with an increase in the LC3 -Ⅱ expression （all P 〈0.05）; compared with groups A and B, the expression of LC3-Ⅰin the group C reduced while the LC3 -Ⅱ expression increased （all P 〈0.05）.Conclusion The inhibition of erlotinib to NSCLC with wild-type EGFR can be

关 键 词：非小细胞肺癌 厄洛替尼 氯喹 表皮生长因子受体 K-RAS基因 细胞增殖 细胞凋亡 细胞自噬 微管相关蛋白轻链3 

分 类 号：R734.2[医药卫生—肿瘤]