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为研究亚硝酸钠对人肝癌SMMC-7721细胞侵袭能力的影响,用0.25mmol·L-1亚硝酸钠孵育SMMC-7721细胞48h,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,Westernblot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48h,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48h,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).0.25mmol·L-1亚硝酸钠处理细胞48h,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.
To investigate the effect of sodium nitrite on invasiveness of human hepatocellular carcinoma SMMC-7721 cells, SMMC-7721 cells were incubated with 0.25 mmol·L-1 sodium nitrite for 48 h and then supernatants of cell culture supernatants were collected.Transplants coated with artificial basement membrane The cells were stained with Coomassie Brilliant Blue and α-tubulin immunofluorescence staining to detect the cytoskeleton.The expression of matrix metalloproteinases (MMPs) was detected by Western blotting.The results showed that SMMC-7721 cells were incubated with sodium nitrite for 48h, (211.5 ± 17.6), which was significantly different from that of the control group (176.3 ± 15.5) (p <0.05). After incubation with sodium nitrite for 48h, the number of cells in the cell culture supernatant + serum- (165.12 ± 7.90), which was significantly higher than that in the negative control group (30.32 ± 11.00) (p <0.05). The cell culture supernatant and the cells containing 10% bovine serum albumin (FBS) (186.73 ± 9.41) cells (170.54 ± 9.60), which was significantly higher than that of 10% FBS positive control group (p <0.05) .0.25mmol·L-1 sodium nitrite treatment of cells 48h, cells Skeletal microfilaments aggregate into stress fibers, extending along the longitudinal axis of the cells, elongating the cell body and polymerizing the tubulin; MM P-2 and MMP-9 protein expression increased.The above results show that sodium nitrite can promote the invasiveness of human hepatoma SMMC-7721 cells by inducing cytoskeletal rearrangements.