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最近发现肿瘤微环境内含有较高水平的亚硝酸盐和氨,但是它们对癌细胞的影响还不清楚。本研究旨在探讨亚硝酸盐和铵盐联合暴露对人肝癌细胞侵袭能力的影响及活性氧(ROS)/鸟氨酸脱羧酶(ODC)途径在其中的作用。经亚硝酸钠、氯化铵、亚硝酸钠和氯化铵混合物处理的SMMC-7721细胞,采用MTT实验检测细胞活力,transwell小室检测细胞侵袭能力,ROS检测试剂盒检测细胞内ROS水平,免疫荧光检测细胞内ODC,Western blot检测基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)和ODC的蛋白表达。结果表明,亚硝酸钠和氯化铵混合物能够增强SMMC-7721细胞侵袭能力,促进细胞内ROS水平增加,促进ODC蛋白表达,ROS清除剂N-乙酰半胱氨酸(NAC)可以逆转上述现象;ODC特异性抑制剂二氟甲基鸟氨酸(DFMO)可以增加细胞内ROS水平,降低细胞侵袭能力。综上所述,亚硝酸钠和氯化铵混合物通过增强ROS/ODC途径,促进人肝癌SMMC-7721细胞侵袭。
The tumor microenvironment has recently been found to contain high levels of nitrite and ammonia, but their effect on cancer cells is unclear. The purpose of this study was to investigate the effect of combined nitrite and ammonium exposure on the invasiveness of human hepatoma cells and the role of reactive oxygen species (ROS) / ornithine decarboxylase (ODC) pathways in them. SMMC-7721 cells treated with a mixture of sodium nitrite, ammonium chloride, sodium nitrite and ammonium chloride were assayed for cell viability by MTT assay, cell invasion assay by transwell chamber, ROS detection by ROS assay kit, immunofluorescence The intracellular ODC was detected and the protein expression of MMP-2, MMP-9 and ODC was detected by Western blot. The results showed that the mixture of sodium nitrite and ammonium chloride could enhance the invasiveness of SMMC-7721 cells, increase the intracellular ROS level and promote the expression of ODC protein, and N-acetyl cysteine (NAC), an ROS scavenger, could reverse the above phenomenon. ODC specific inhibitor difluoromethylornithine (DFMO) can increase intracellular ROS levels and reduce cell invasiveness. In summary, the mixture of sodium nitrite and ammonium chloride enhances the invasion of human hepatocellular carcinoma SMMC-7721 cells by enhancing the ROS / ODC pathway.