FK866对非小细胞肺癌细胞迁移的影响  

Effects of FK866 on migration of A549 cells and related mechanism

作　　者：谢娴 徐小方[2,3] 王琪[2] 卢韵碧[1] 吴明[2] 张纬萍[1] XIE Xian;XU Xiaofang;WANG Qi;LU Yunbi;WU Ming;ZHANG Weiping(Department of Pharmacology, Zhejiang University School of Medicine, Hangzhou 310058, China;Department of Thoracic Surgery, Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310009, China;Department of Thoracic Surgery, Zhejiang Cancer Hospital, Hangzhou 310022, China)

机构地区：[1]浙江大学医学院药理学系,浙江杭州310058 [2]浙江大学医学院附属第二医院胸外科,浙江杭州310009 [3]浙江省肿瘤医院胸外科,浙江杭州310022

出　　处：《浙江大学学报（医学版）》2018年第1期1-9,共9页Journal of Zhejiang University(Medical Sciences)

基　　金：浙江省自然科学基金(LY16H010004;LY18H170001;LQ17H010002);国家自然科学基金(81573400);国家科技支撑计划(2015BAI13B02);浙江省公共技术应用研究(2016F82G2010036)

摘　　要：目的:探讨低浓度烟酰胺磷酸核糖基转移酶(NAMPT)抑制剂FK866对人非小细胞肺癌细胞株(A549细胞)迁移的影响及作用机制。方法:MTT法检测不同浓度FK866对A549细胞增殖的影响;划痕实验检测1.0 nmol/L和10.0 nmol/L FK866对A549细胞迁移的影响;实时定量RT-PCR检测上皮间质转化相关蛋白上皮钙黏素和波形蛋白mRNA的表达量;蛋白质印迹法检测细胞外调节蛋白激酶1/2(ERK1/2)和磷酸化ERK1/2蛋白的表达量。结果:FK866作用时间越长、浓度越高,对A549细胞增殖的抑制作用越明显。FK866作用72 h时的半抑制浓度(IC50)为9.55 nmol/L。以1.0、10.0 nmol/L的FK866预处理细胞48 h,划痕后继续给药48 h,两种浓度的FK866均能抑制A549细胞迁移;如不进行预处理,仅10.0 nmol/L浓度的FK866对划痕愈合有一定的抑制作用;1.0 nmol/L的FK866处理细胞72 h可上调上皮钙黏素和波形蛋白mRNA表达,并激活ERK1/2;以1.0 mmol/L的烟酰胺单核苷酸(NMN)或10.0μmol/L的ERK1/2抑制剂U0126预处理,能够逆转FK866引起的上皮钙黏素和波形蛋白表达上调。结论:低浓度FK866能够通过减少细胞内烟酰胺腺嘌呤二核苷酸和激活ERK1/2,增加上皮钙黏素表达,进而抑制细胞迁移,对肿瘤转移可能有一定的抑制作用;但其同时促进了波形蛋白的表达,不利于肿瘤的治疗。Objective： To investigate the effect of nicotinamide phosphoribosyltransferase（NAMPT） inhibitor FK866 on the migration of human non-small cell cancer A549 cells and related mechanism. Methods： The inhibition effect of FK866 on A549 cells was tested by MTT assay. A549 cells were treated with 1. 0 and 10. 0 nmol/L FK866,and the cell migration was evaluated by modified wound scratch assay. The mRNA expression of E-cadherin and vimentin was detected by real-time RT-PCR,and the expression of ERK1/2 and p ERK1/2 was determined by Western blotting. Results：FK866 inhibited the proliferation of A549 cells in a time-and concentration-dependent manner; after treatment for 72 h, the IC50 of FK866 was 9. 55 nmol/L. When1. 0 nmol/L or 10. 0 nmol/L FK866 was continuously applied 48 h before and 48 h after a scratch was made in wound scratch assay, the migration of A549 cells was significantly inhibited. However,when the FK866 was applied only 48 h after the scratch,the migration of A549 cells was inhibited by 10. 0 nmol/L but not by1. 0 nmol/L FK866. The mRNA expression of E-cadherin and vimentin, and the activated ERK1/2 were significantly increased after 1. 0 nmol/L FK866 treatment for72 h. The pretreatment with nicotinamide adenine dinucleotide（ NAD） precursor nicotinamide mononucleotide（1. 0 mmol/L） or ERK1/2 inhibitor U0126（10. 0 μmol/L）reversed the up-regulation of E-cadherin and vimentin expression induced by FK866.Conclusions： Low concentration of FK866 decreases the migration of A549 cells through the inhibition of NAD level, activation of ERK1/2 and up-regulation of E-cadherin expression. However, it also up-regulates the expression of vimentin,indicating that it may have dual effects on the migration of tumor cells.

关 键 词：烟酰胺磷酸核糖基转移酶/拮抗剂和抑制剂 烟酰胺磷酸核糖基转移酶/药理学 癌 非小细胞肺/病理学 钙黏着糖蛋白类/代谢 波形蛋白/代谢 细胞运动 肿瘤细胞 培养的/药物作用 

分 类 号：R961[医药卫生—药理学]