下调IGF-1R对人肺腺癌A549细胞吉非替尼耐药性的影响  

Effect of down-regulating IGF-1R on drug resistance of human lung cancer A549 cells to gefitinib

作　　者：刘俊[1] 宁兰兰[1] 赵子文[1] 马为[1] 王汉平[1] 徐邦牢[1] LIU Jun;NING Lanlan;ZHAO Ziwen;MA Wei;WANG Hanping;XU Banglao(Guangzhou First People's Hospital of Guangzhou Medical University, Guangzhou 510580, China)

机构地区：[1]广州医科大学附属广州市第一人民医院,广州510180

出　　处：《山东医药》2019年第7期41-44,共4页Shandong Medical Journal

基　　金：广州市医药卫生科技项目(20151A010003)

摘　　要：目的探讨下调胰岛素样生长因子1受体(IGF-1R)对人肺腺癌A549细胞吉非替尼耐药性的影响。方法取人肺腺癌细胞株A549为亲本细胞体外培养;采用低浓度吉非替尼逐步加量诱导法,建立吉非替尼耐药细胞株A549/GR(阳性对照组);随后构建慢病毒IGF-1R-siRNA表达载体和NC-siRNA表达载体,并用脂质体法分别转染亲本细胞株,将其设为实验组和阴性对照组。用real-time RCR法、Western blotting法分别检测实验组和阴性对照组中的IGF-1R的mRNA和蛋白。采用CCK-8比色法测算实验组、阴性对照组吉非替尼培养12、24、48 h时的细胞活性。结果实验组细胞IGF-1R的mRNA、蛋白相对表达量均比阴性对照组下调(P均<0.05)。实验组吉非替尼培养24、48 h时的细胞活力较阴性对照组低(P均<0.01)。结论下调IGF-1R可减弱人肺腺癌A549细胞对吉非替尼的耐药性。Objective To analyze the effect of down-regulating insulin-like growth factor-1 receptor (IGF-1R) on the drug resistance of human lung cancer A549 cells to gefitinib.Methods We used human lung cancer cell line A549 as the parent cells in vitro, and established the gefitinib-resistant cell line A549/GR by using gradually incremental induction of gefitinib (positive control group). We constructed IGF-1R-siRNA and NC-siRNA targeting random sequence lentiviral vectors, and transfected two vectors into A549 cells by using Lipo-2000 to establish the experimental group and negative control group, respectively. The expression levels of mRNA and protein were detected by RT-qPCR and Western blotting in the cells of the experimental group and negative control group. In addition, we calculated the cell viability at 12, 24, and 48 h after gefitinib treatment in the experimental group and negative control group by CCK-8 colorimetric method. Results After transfection, the mRNA and protein levels of IGF-1R in the experimental group were significantly down-regulated as compared with those in the negative control group (both P <0.05). The cell viability of the experimental group was significantly lower than that of the negative control group at 24 and 48 h after gefitinib treatment (both P <0.01). Conclusion Down-regulation of IGF-1R decreases the drug resistance of human lung adenocarcinoma A549 cells to gefitinib.

关 键 词：胰岛素样生长因子1受体 肺肿瘤 吉非替尼 细胞耐药 

分 类 号：R734.2[医药卫生—肿瘤]