目的:探讨姜黄素抗胶质瘤作用的分子机制。方法:(一)细胞实验：(1)取对数生长期的U251MG、SHG-44细胞，分别分为姜黄素组与对照组、阴性对照小干扰RNA(siRNA)组与H19 siRNA组、阴性对照siRNA+姜黄素组与H19 siRNA+姜黄素组、H19 siRNA+阴性对照抑制物组与H19 siRNA+miR-491-5p抑制物组、H19 siRNA+阴性对照抑制物+姜黄素组与H19 siRNA+miR-491-5p抑制物+姜黄素组、miR-491-5p模拟物+空白质粒+姜黄素组与miR-491-5p模拟物+同源盒基因A9(HOXA9)过表达质粒+姜黄素组，各组细胞分别予10 μmol/L姜黄素或阴性对照siRNA、H19 siRNA转染或miRNA抑制物、miR-491-5p抑制物共转染或miR-491-5p模拟物+空白质粒、miR-491-5p模拟物+HOXA9过表达质粒共转染等不同处理。采用实时荧光定量PCR(qRT-PCR)法检测各组细胞中n H19、n miR-491-5p、n HOXA9 mRNA的表达水平，采用细胞计数试剂(CCK)-8法检测各组细胞的增殖率，采用流式细胞术检测各组细胞的凋亡率，采用平板克隆法检测各组细胞的克隆形成数，采用Transwell小室实验检测各组细胞的迁移情况，采用Western blotting法检测各组细胞中HOXA9蛋白的表达水平。(2)取对数生长期的293T细胞，分别分为阴性对照模拟物+野生型H19组与miR-491-5p模拟物+野生型H19组、阴性对照模拟物+野生型HOXA9 3'-UTR组与miR-491-5p模拟物+野生型HOXA9 3'-UTR组，各组细胞分别予阴性对照miRNA模拟物、miR-491-5p模拟物与野生型H19、野生型HOXA9 3'-UTR质粒载体共转染等不同处理。采用双荧光素酶报告实验检测各组细胞的荧光素酶活性。(二)病例标本实验：收集南阳市中心医院神经外科自2017年5月至2019年5月手术切除且经病理检查确诊为胶质瘤的30例组织标本(胶质瘤组)及同期行内减压术获得的30例正常脑组织标本(正常组)，采用qRT-PCR法检测各组标本中n H19、n miR-491-5p、n HOXA9 mRNA的表达水平，采用Western blotting法检测各组标本中HOXA9蛋白的表达水平。(三)裸鼠实验：将24只裸鼠按随机数字表法分为阴性对照shRNA组、H19 shRNA组、阴性对照shRNA+姜黄素、H19 shRNA+姜黄素组，每组6只，分别腹腔注射稳定转染阴性对照shRNA、H19 shRNA的U251MG细胞以及次日注射60 mg/kg剂量姜黄素。分别于饲养第7、11、15、19、23、27天时测量各组大鼠的肿瘤体积，并采用qRT-PCR法检测肿瘤组织中n H19、n miR-491-5p、n HOXA9 mRNA的表达水平，采用Western blotting法检测HOXA9蛋白的表达水平。n 结果:(一)细胞实验：(1)姜黄素组与对照组相比、H19 siRNA组与阴性对照siRNA组相比，前者U251MG、SHG-44细胞中n H19、n HOXA9 mRNA和HOXA9蛋白的表达水平明显降低，n miR-491-5p mRNA的表达水平明显升高，差异均有统计学意义(n P<0.05)。H19 siRNA+miR-491-5p抑制物组与H19 siRNA+阴性对照抑制物组相比，前者U251MG、SHG-44细胞中HOXA9 mRNA和蛋白的表达水平明显升高，差异均有统计学意义(n P<0.05)。姜黄素组与对照组相比、H19 siRNA组与阴性对照siRNA组相比、H19 siRNA+姜黄素组与阴性对照siRNA+姜黄素组相比，前者U251MG、SHG-44细胞培养72 h时的细胞增殖率明显降低，细胞凋亡率明显升高，细胞克隆形成数明显降低，细胞迁移数明显降低，差异均有统计学意义(n P<0.05)。H19 siRNA+miR-491-5p抑制物+姜黄素组与H19 siRNA+阴性对照抑制物+姜黄素组相比、miR-491-5p模拟物+HOXA9过表达质粒+姜黄素组与miR-491-5p模拟物+空白质粒+姜黄素组相比，前者U251MG、SHG-44细胞培养72 h时的细胞增殖率明显升高，细胞凋亡率明显降低，细胞克隆形成数明显升高，细胞迁移数明显升高，差异均有统计学意义(n P<0.05)。(2)miR-491-5p模拟物+野生型H19组与阴性对照模拟物+野生型H19组相比、miR-491-5p模拟物+野生型HOXA9 3'-UTR组与阴性对照模拟物+野生型HOXA9 3'-UTR组相比，前者细胞荧光素酶活性明显降低，差异均有统计学意义(n P<0.05)。(二)病例标本实验：胶质瘤组与正常组相比，前者标本中n H19、n HOXA9 mRNA和HOXA9蛋白的表达水平明显升高，n miR-491-5p mRNA的表达水平明显降低，差异均有统计学意义(n P<0.05)。(三)裸鼠实验：饲养第27天时，H19 shRNA组与阴性对照shRNA组相比、H19 shRNA+姜黄素组与阴性对照shRNA+姜黄素组相比，前者肿瘤体积明显降低，肿瘤组织中n miR-491-5p mRNA的表达水平明显升高，n H19 mRNA、HOXA9 mRNA和蛋白的表达水平明显降低，差异均有统计学意义(n P<0.05)。n 结论:姜黄素通过长链非编码RNA H19/miR-491-5p/HOXA9轴抑制胶质瘤细胞的增殖、迁移及促进细胞凋亡。“,”Objective:To investigate the molecular mechanism of antiglioma effect of curcumin.Methods:Cell experiment: (1) U251MG and SHG-44 cells at logarithmic growth phase were treated with 10 μmol/L curcumin (curcumin group) or same volume of dimethyl sulfoxide solution (control group); cells were transfected with negative control small interfering RNA (siRNA) and long non-coding RNA (lncRNA) H19 siRNA (negative control siRNA group and H19 siRNA group); cells were transfected with negative control siRNA and H19 siRNA, respectively, and then, they were treated with 10 μmol/L curcumin (negative control siRNA+curcumin group and H19 siRNA+curcumin group); the H19 siRNA was co-transfected with negative control miR inhibitor or miR-491-5p inhibitor into these cells (H19 siRNA+negative control inhibitor group and H19 siRNA+miR-4915p inhibitor group); H19 siRNA+negative control miR inhibitor or H19 siRNA+miR-491-5p inhibitor were co-transfected into the cells, and then, they were treated with 10 μmol/L curcumin (H19 siRNA+negative control inhibitor+curcumin group and H19 siRNA+miR-491-5p inhibitor+curcumin group); the cells were co-transfected with miR-491-5p mimic+blank plasmid or miR-491-5p mimic+HOXA9 overexpression plasmid, and then they were treated with 10 μmol/L curcumin (miR-491-5p mimic+blank plasmid+curcumin group and miR-491-5p mimic+HOXA9 overexpression plasmid+curcumin group); real-time fluorescent quantitative PCR (qRT-PCR) was used to detect the mRNA expressions of n H19, n miR-491-5p, and n HOXA9; CCK-8 assay was used to detect the cell proliferation; flow cytometry was used to detect the cell apoptosis; plate cloning method was employed to detect the number of cell clone formation; Transwell assay was used to detect the cell migration; and the HOXA9 protein expression was measured by Western blotting. (2) The 293T cells at the logarithmic growth phase were chosen; the negative control miRNA mimics or miR-491-5p mimics combined with wild-type H19, wild-type HOXA9 3'-UTR plasmid vectors were co-transfected into the cells, respectively (negative control mimic+wild type H19 group and miR-491-5p mimic+wild type H19 group, negative control mimic+wild type HOXA9 3'-UTR group and miR-491-5p mimic+wild type HOXA9 3'-UTR group); the luciferase activity was detected by dual luciferase reporter experiment. (3) Thirty specimens from glioma patients (glioma group) underwent surgical resection and pathologically confirmed in our hospital from May 2017 to May 2019 and 30 normal brain tissue specimens obtained during decompression (normal group) at the same period were chosen; the mRNA expressions ofn H19, n miR-491-5p, and n HOXA9 were detected by qRT-PCR, and the HOXA9 protein expression level in these specimens was detected by Western blotting. (4) Twenty-four nude mice were randomly divided into negative control short hairpin RNA (shRNA) group, H19 shRNA group, negative control shRNA+curcumin group, and H19 shRNA+curcumin group (n n=6); U251MG cells stably transfected with negative control shRNA or H19 shRNA were intraperitoneally injected, respectively, into the mice; and 60 mg/kg curcumin was injected on the next d; the tumor volume was measured on the 7n th, 11n th, 15n th, 19n th, 23n rd, and 27n th d of rearing; and the n H19, n miR-491-5p and n HOXA9 mRNA expressions in the tumor tissues were detected by qRT-PCR; the HOXA9 protein expression was detected by Western blotting.n Results:(1) When curcumin group comparing with control group, and H19 siRNA group comparing with negative control siRNA group, U251MG and SHG-44 cells had significantly decreased miR-491-5p mRNA and protein expressions, and significantly increased n miR-491-5p mRNA expression (n P<0.05); as compared with that in the H19 siRNA+negative control inhibitor group, the HOXA9 mRNA and protein expressions in U251MG and SHG-44 cells of H19 siRNA+miR-491-5p inhibitor group were significantly higher (n P<0.05). When curcumin group comparing with control group, H19 siRNA group comparing with negative control siRNA group, H19 siRNA+curcumin group comparing with negative control siRNA+curcumin group, the U251MG and SHG-44 cells after 72 h of culture had significantly decreased cell proliferation rate, significantly increased apoptosis rate, significantly reduced number of cell clone formation, and significantly reduced cell migration number (n P<0.05). When H19 siRNA+miR-491-5p inhibitor+curcumin group comparing with H19 siRNA+negative control inhibitor+curcumin group, miR-491-5p mimic+HOXA9 overexpression plasmid+curcumin group comparing with miR-491-5p mimic+blank plasmid+curcumin group, the U251MG and SHG-44 cells after 72 h of culture had significantly increased cell proliferation rate, significantly reduced apoptosis rate, significantly increased number of cell clone formation, and significantly increased cell migration number (n P<0.05). (2) When miR-491-5p mimic+wild-type H19 group comparing with negative control mimic+wild-type H19 group, miR-491-5p mimic+wild-type HOXA9 3'-UTR group comparing with negative control mimic+wild-type HOXA9 3'-UTR group, the cell luciferase activity was significantly reduced (n P<0.05). (3) As compared with those in the normal group, then H19 and n HOXA9 mRNA expressions and HOXA9 protein expression in the glioma group were significantly increased, and the n miR-491-5p mRNA expression was significantly reduced (n P<0.05). (4) On the 27n th d of rearing, when H19 shRNA group comparing with negative control shRNA group, and H19 shRNA+curcumin group comparing with negative control shRNA+curcumin group, the tumor volume was significantly reduced, the n miR-491-5p mRNA expression in the tumor tissues was significantly increased, and the n H19 mRNA, HOXA9 mRNA and protein expressions were significantly reduced (n P<0.05).n Conclusion:Curcumin may inhibit the cell proliferation and migration and promote the apoptosis of glioma cells through lncRNA H19/miR-491-5p/HOXA9 axis.