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目的探讨聚肌苷酸胞苷酸(PolyI:C)对体外培养的人肝癌细胞SMMC-7721生长的增殖抑制和凋亡诱导作用,并探讨其作用机制。方法肝癌SMMC-7721细胞经不同剂量PolyI:C作用后,CCK-8法检测细胞增殖抑制,流式细胞仪检测细胞凋亡及细胞周期变化,SYBR-Green荧光定量PCR检测TLR3、TRIF、IFN-βmRNA的表达。结果PolyI:C高剂量组的生长抑制率最高,高、中、低三个剂量组间比较差异有显著性(P<0.05)。PolyI:C同一剂量组生长抑制率72h最高,48h次之,24h时最低,每组不同时间点比较差异有显著性(P<0.05)。流式细胞仪检测结果显示PolyI:C可诱导SMMC-7721细胞凋亡,随浓度增加和时间的延长凋亡率逐渐升高(P<0.05)。PolyI:C组细胞处于G1期的百分率明显高于对照组。TLR3、TRIF、IFN-βmRNA的表达均增高,三组间比较差异有显著性(P<0.05)。结论PolyI:C对体外培养的SMMC-7721有显著的增殖抑制作用,呈剂量依赖性和时间依赖性,其机制可能与细胞周期阻滞,TLR3通路激活,诱导产生IFN-β,诱导细胞凋亡有关。
Objective To investigate the effects of poly I (C) on the proliferation and apoptosis of human hepatocellular carcinoma cell line SMMC-7721 in vitro and to explore its possible mechanism. Methods The proliferation of SMMC-7721 cells were detected by CCK-8 assay, the apoptosis and cell cycle were detected by flow cytometry. The expressions of TLR3, TRIF and IFN-γ were detected by SYBR-Green fluorescence quantitative PCR. β mRNA expression. Results PolyI: C high-dose group had the highest growth inhibition rate, high, medium and low dose groups showed significant difference (P <0.05). PolyI: C in the same dose group had the highest growth inhibition rate at 72h, 48h, and lowest at 24h, with significant difference at different time points (P <0.05). Flow cytometry results showed that PolyI: C could induce apoptosis of SMMC-7721 cells, and the apoptosis rate increased gradually with the increase of concentration and time (P <0.05). PolyI: The percentage of cells in G1 phase in group C was significantly higher than that in control group. The expressions of TLR3, TRIF and IFN-βmRNA were all increased in the three groups (P <0.05). Conclusion PolyI: C can significantly inhibit the proliferation of SMMC-7721 cells in a dose-and time-dependent manner. The mechanism may be related to cell cycle arrest, activation of TLR3 pathway, induction of IFN-β production, and induction of apoptosis related.