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目的:观察外源锌对原发性肝细胞癌(HCC)BEL-7404细胞生物学行为的影响。方法:应用TSQ锌离子荧光探针、MTT法、DNA倍体法、吖啶橙/溴乙啶双荧光染色法和Transwell小室法分别检测不同浓度硫酸锌刺激下BEL-7404细胞内锌离子含量、细胞活力、细胞周期、细胞凋亡及细胞迁移和侵袭能力的变化。应用real-time PCR和Western blot法分别检测不同浓度锌离子对BEL-7404细胞白蛋白的mRNA和蛋白表达的影响。结果:随着培养环境中锌离子浓度的升高,BEL-7404细胞内的锌离子含量增加,细胞的存活率和细胞迁移与侵袭能力降低,凋亡率升高(P<0.05);G_0/G_1期细胞比例降低,G_2/M期细胞比例升高(P<0.05);细胞白蛋白的mRNA和蛋白表达量增加(P<0.05)。结论:外源性给予锌离子可抑制HCC细胞的活力、迁移与侵袭能力,诱导细胞凋亡,阻滞细胞周期在G_2/M期,并可能降低细胞的恶性表型。
Objective: To observe the effect of exogenous zinc on the biological behavior of primary hepatocellular carcinoma (HCC) BEL-7404 cells. METHODS: The zinc ion content in BEL-7404 cells stimulated by different concentrations of zinc sulfate was detected by TSQ zinc ion fluorescence probe, MTT assay, DNA ploidy method, acridine orange/ethidium bromide staining method and Transwell chamber method. Cell viability, cell cycle, apoptosis, and changes in cell migration and invasiveness. Real-time PCR and Western blot were used to detect the effects of different concentrations of zinc on albumin mRNA and protein expression in BEL-7404 cells. Results: With the increase of zinc ion concentration in the culture environment, the zinc ion content in BEL-7404 cells increased, and the cell survival rate, cell migration and invasion ability decreased, and the apoptosis rate increased (P<0.05); G_0/ The proportion of cells in G 1 phase decreased, the proportion of cells in G 2 /M phase increased (P<0.05), and the expression of cellular albumin mRNA and protein increased (P<0.05). CONCLUSION: Exogenous administration of zinc ions can inhibit the viability, migration and invasion of HCC cells, induce apoptosis, arrest the cell cycle in G 2 /M phase, and may reduce the malignant phenotype of HCC cells.