盐酸埃克替尼对非小细胞肺癌细胞增殖、凋亡的影响及其机制探讨  

Effects of icotinib hydrochloride on proliferation and apoptosis of non-small-cell lung cancer cells

作　　者：王林梅[1] 刘剑波[1] 邵润霞[1] 齐景宪[1] 尚学琴 

机构地区：[1]郑州大学第二附属医院,郑州450014 [2]昆明市第二人民医院

出　　处：《山东医药》2016年第38期9-12,共4页Shandong Medical Journal

基　　金：河南省医学科技攻关计划项目(201403899)

摘　　要：目的观察盐酸埃克替尼对非小细胞肺癌PC-9细胞增殖、凋亡的影响,并探讨其机制。方法取对数生长期的PC-9细胞,以40μmol/L盐酸埃克替尼培养0、2、4、8、12、24 h,以0、20、40、60、80μmol/L的盐酸埃克替尼处理12 h,采用MTT法检测细胞增殖活性;将对数生长期的PC-9细胞分为给药组和对照组,分别以40μmol/L盐酸埃克替尼、DMSO处理8 h,用流式细胞仪检测凋亡细胞并计算凋亡率,Western blot法检测JAK2、p-JAK2、信号转导与激活因子3(STAT3)、p-STAT3和血管内皮生长因子(VEGF)蛋白,细胞免疫荧光技术观察并计算STAT3入核细胞百分率。结果随着盐酸埃克替尼浓度的增加及作用时间的延长,PC-9细胞增殖活性逐渐降低,40、60、80μmol/L与0μmol/L-比较,8、12、24 h与0 h比较,P均<0.05。给药组PC-9细胞凋亡率3.70%±0.39%,明显高于对照组的1.40%±0.21%,P<0.05。与对照组比较,给药组p-JAK2、p-STAT3相对表达量均减少(P均<0.05),VEGF蛋白相对表达量亦减少(P<0.01)。给药组STAT3入核细胞百分率为36.48%±2.12%,低于对照组的86.56%±1.82%(P<0.01)。结论盐酸埃克替尼可抑制非小细胞肺癌细胞增殖,促进细胞凋亡;其可能通过抑制JAK2-STAT3信号通路活性及STAT3入核,并下调VEGF蛋白表达发挥作用。Objective To observe the effect of icotinib hydrochloride on the proliferation and apoptosis of non-small- cell lung cancer （NSCLC） PC-9 cells and investigate its underlying mechanism. Methods PC-9 cells in the logarithmic phase was treated with 40 μmol/L icotinib hydrochloride for 0, 2, 4, 8, 12 and 24 h, followed by 0, 20, 40, 60 and 80 μmol/L icotinib hydrochloride of 12 h. MTT was used to detect the proliferation activity of PC-9 cells. PC-9 cells were divided into the experimental group （which was treated with 40 mmol/L ieotinib hydroehloride for 8 h） and control group （which was treated with DMSO for 8 h）. The apoptosis rate of PC-9 cells was determined by flow cytometry. The protein expression of JAK2, p-JAK2, signal transducer and activator oftranscription 3 （ STAT3 ）, p-STAT3 and vascular endothelial growth factor （VEGF） was detected by Western blotting. The translocation of STAT3 and its transcation on VEGF was re- spectively measured by cell immunochemistry. Results With the increasing concentrations of icotinib hydrochloride or with the extension of time, the proliferation activity of PC-9 cells gradually reduced. Significant difference was found between 40, 60, 80 μmol/L and 0 μmol/L, between 8, 12, 24 h and 0 h （ all P 〈 0.05）. The apoptosis rate of PC-9 cells in the experimental group was 3.7% ±0.39%, significantly higher than that of the control group （ 1.4%±0.21% ）, P 〈 0.05. Compared with the control group, the relative expression of p-JAK2 and p-STAT3 was reduced （ P 〈 0.05 ） as well as the relative expression of VEGF protein in the experimental group （ P 〈 0.01）. The cell percentage of STAT3 in nuclear was 36.48%±2.12%, which was lower than that of the control group （86.56% ± 1.82% ）, （P 〈0.01 ）. Conclusion Icotinib hydroehloride could inhibit the cell proliferation, promote the apoptosis of PC-9 cells through inhibiting the activity of JAK2-STAT3 signaling pathway and cell percentage of STAT3 in nuclear as well as down-regulating V

关 键 词：非小细胞肺癌 PC-9细胞 盐酸埃克替尼 细胞增殖 细胞凋亡 信号通路 血管内皮生长因子 

分 类 号：R915[医药卫生—微生物与生化药学]