• 关键词：亚硝酸钠  人肝癌SMMC-7721细胞株  侵袭  细胞骨架

• 基金项目：国家重大环保公益项目专项(No.200809115 );省部共建河南大学科研项目(No.SBGJ090702)

• 闫文义
• 河南大学医学院环境医学研究所,开封 475004

• 皇甫超申
• 1. 河南大学医学院环境医学研究所,开封 475004;
  2. 河南大学环境与健康工程中心,开封 475004

• 史齐
• 河南大学医学院环境医学研究所,开封 475004

• 李延红
• 河南大学医学院环境医学研究所,开封 475004

• 石贞玉
• 河南大学医学院环境医学研究所,开封 475004

• 厉永强
• 河南大学医学院环境医学研究所,开封 475004

• 刘彬
• 河南大学医学院环境医学研究所,开封 475004

• 摘要：为研究亚硝酸钠对人肝癌SMMC-7721细胞侵袭能力的影响,用0.25 mmol · L^-1 亚硝酸钠孵育SMMC-7721细胞48 h,然后收集细胞培养液上清.用人工基底膜包被的Transwell小室记录细胞侵袭能力,考马斯亮蓝及α-微管蛋白免疫荧光染色观察细胞骨架,Western blot检测细胞基质金属蛋白酶(MMPs)表达.结果显示,用亚硝酸钠孵育SMMC-7721细胞48 h,穿膜细胞为(211.5±17.6)个,与对照组(176.3±15.5)个有显著性差异(p﹤0.05);用亚硝酸钠孵育48 h,细胞培养液上清+无血清培养液组细胞穿膜数为(165.12±7.90)个,较无血清培养液阴性对照组(30.32±11.00)个显著增多(p﹤0.05);细胞培养液上清+含10%牛血清白蛋白(FBS)组细胞穿膜数为(186.73±9.41)个,较单纯含10%FBS阳性对照组细胞穿膜数(170.54±9.60)个也明显增多(p﹤0.05).0.25 mmol · L^-1 亚硝酸钠处理细胞48 h,细胞骨架微丝聚合成应力纤维,沿细胞纵轴延伸,使细胞胞体伸长,微管蛋白聚合;MMP-2 和MMP-9蛋白表达增加.上述结果表明,亚硝酸钠通过诱导细胞骨架重排,促进人肝癌SMMC-7721细胞侵袭能力.

• Abstract：The aim of this study was to investigate the effect of sodium nitrite on the invasion of human hepatoma cell line SMMC-7721.The cell supernatants were collected from SMMC-7721 cells treated with 0.25 mmol.L^-1 NaNO₂ for 48 h.The tumor cell invasion was investigated with matrigel-coated transwell chamber assay.The change of cytoskeleton was observed by staining with Coomassie blue or immunofluorescence.The expression of matrix metalloproteinases (MMPs) protein in SMMC-7721 cells was detected by western blot.After treating SMMC-7721 cells with 0.25 mmol · L^-1 NaNO₂ for 48 h, the matrigel-coated invasion chamber assay was conducted.The results showed that the number of migrated cells was 211.5±17.6, and that of the control group was 176.3±15.5, indicating that NaNO₂ could significantly increase cell migration (p<0.05).More cells invaded the ECM-coated filter in the group of serum-free media mixed with cell supernatants (165.12±7.90, p<0.05), compared with those in the serum-free, media-only group (negative control group, 30.32±11.00).The number of migrated cells in the group of media containing 10% fetal bovine serum (FBS) and cell supernatants (186.73±9.41) were also higher than those in the group of 10% FBS only (positive control, 170.54±9.60, p<0.05).After treatment with 0.25 mmol · L^-1 NaNO₂ for 48 h, the cellular cytoskeleton F-actin fibers distributing in the perimeter aggregated into intermediate filaments (stress fiber) and elongated along the vertical axis, resulting in elongation of the cellular body, the polymerization of tubulin and the over-expression of MMP-2 and MMP-9 protein.The results demonstrated that NaNO₂ could induce cytoskeletal rearrangement and promote the capability of invasion in SMMC-7721 cells.

• Key words：sodium nitrite  human hepatoma SMMC-7721 cells  invasion  cytoskeleton

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