托芬那酸通过下调LOXL2表达抑制人胃癌SGC-7901细胞侵袭  

Suppressing invasion of human gastric cancer SGC-7901 cells by tolfenamic acid through inhibition of LOXL2

作　　者：程琦[1] 徐松涛[1] 饶淑梅[1] 马永超[1] CHENG Qi;XU Song-Tao;RAO Shu-Mei;MA Yong-Chao(Luohe Medical College,Luohe 462002,China)

机构地区：[1]漯河医学高等专科学校,漯河462002

出　　处：《中国免疫学杂志》2020年第21期2624-2627,2634,共5页Chinese Journal of Immunology

基　　金：漯河医学高等学校创业创新发展能力提升工程项目(2019-LYZTD005)资助。

摘　　要：目的:观察托芬那酸(TA)对人胃癌SGC-7901细胞侵袭能力的影响,并探讨其作用机制。方法:用不同剂量的TA(0、3、6、12、24、48、96μmol/L)处理SGC-7901细胞24 h或48 h,CCK-8法检测细胞活力,脂质体介导shRNA转染靶向沉默LOXL2基因,Transwell检测侵袭细胞数,RT-PCR检测LOXL2的mRNA水平,Western blot检测SP1、LOXL2、Snail、E-cad、MMP9蛋白水平。结果:TA可剂量依赖性地抑制SGC-7901细胞活力(P<0.05),24 h和48 h的IC50分别为78.06μmol/L和50.72μmol/L;与对照组相比,0、3、6和12μmol/L TA作用24 h后,SGC-7901细胞侵袭数显著减少(P<0.05),Snail、MMP9、SP1和LOXL2蛋白水平均显著降低(P<0.05),E-cad蛋白水平显著升高(P<0.05),LOXL2的mRNA水平显著降低(P<0.05),且呈现剂量依赖性;敲除LOXL2基因可抑制SGC-7901细胞侵袭(P<0.05),下调Snail和MMP9(P<0.05)、上调E-cad蛋白水平(P<0.05)。结论:TA可能通过下调SP1和LOXL2蛋白表达,进而下调Snail和MMP9、上调E-cad蛋白水平,最终抑制人胃癌SGC-7901细胞侵袭。Objective:To investigate effects of tolfenamic acid(TA)on invasion in human gastric cancer SGC-7901 cells and explore its underlying mechanism.Methods:SGC-7901 cells were treated with TA(0,3,6,12,24,48,96μmol/L)for 24 h and 48 h,cell viability was detected by CCK-8 assay,shRNA plasmid targeting LOXL2 was transfected into SGC-7901 cells by means of liposome assay,Transwell assay was used to determine ability of invasion.The mRNA expression level of LOXL2 were measured by RT-PCR,Western blot was performed to analyze protein levels of SP1,LOXL2,Snail,E-cad and MMP9.Results:TA remarkably reduced cell viability of SGC-7901 cells in a dose-dependent manner(P<0.05),and the IC50 during 24 h and 48 h was 78.06μmol/L and 50.72μmol/L respectively.Compared with control,after exposed to TA(0,3,6 and 12μmol/L)for 24 h,numbers of invasive cells were significantly reduced(P<0.05),protein levels of SP1,LOXL2,Snail and MMP9 were down-regulated(P<0.05),protein level of E-cad was up-regulated(P<0.05),mRNA level of LOXL2 was decreased(P<0.05),in a dose-dependent manner.LOXL2 knockdown also inhibited invasion of SGC-7901 cells(P<0.05),and down-regulated protein levels of Snail and MMP9(P<0.05),and up-regulated protein level of E-cad(P<0.05).Conclusion:TA down-regulates protein levels of Snail and MMP9,and up-regulates protein level of E-cad most likely through inhibiting expression of SP1 and LOXL2 protein,and ultimately lead to a decrease of invasive ability in human gastric SGC-7901 cells.

关 键 词：托芬那酸 胃癌 SGC-7901细胞 侵袭 赖氨酰氧化酶样蛋白2 

分 类 号：R735.2[医药卫生—肿瘤]