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目的观察盐酸青藤碱对人肝癌细胞SMMC-7721凋亡的影响,及其IL-1β、TNF-α及C/EBPβ水平的变化,探讨防治肝癌作用的可能机制。方法体外培养人肝癌细胞SMMC-7721,经不同浓度盐酸青藤碱作用后,采用MTT法检测细胞增殖;Annexin V-FITC/PI双标结合流式细胞术检测细胞的凋亡;ELISA法检测培养液中IL-1β、TNF-α的水平;Western blot技术检测盐酸青藤碱作用于人肝癌细胞后C/EBPβ的表达水平。结果盐酸青藤碱对人肝癌细胞的生长有明显的抑制作用,并呈剂量、时间依赖性,不同浓度组之间差异均有统计学意义(P<0.05);不同浓度(0、0.5、1和2mmol/L)的盐酸青藤碱处理肝癌细胞48h后,各组中凋亡细胞率分别为(8.31±0.81)%、(12.62±1.13)%;(27.43±0.97)%;(51.74±0.85)%,实验组与对照组之间差异有统计学意义(P<0.05);同时发现,与阴性对照组比较,盐酸青藤碱作用组SMMC-7721细胞上清液中IL-1β,TNF-α水平均明显下降,而C/EBPβ的表达水平显著增加(P<0.05)。结论盐酸青藤碱可抑制人肝癌细胞SMMC-7721的细胞增殖,促进其凋亡,此作用可能与其抑制转录因子C/EBPβ,炎症因子如TNF-α、IL-1β的表达,进而通过破坏肿瘤微环境作用有关。
Objective To observe the effect of sinomenine hydrochloride on the apoptosis of human hepatocellular carcinoma cell line SMMC-7721 and the changes of IL-1β, TNF-α and C / EBPβ levels and to explore the possible mechanism of prevention and treatment of hepatocellular carcinoma. Methods Human hepatocellular carcinoma cell line SMMC-7721 was cultured in vitro. MTT assay was used to detect the proliferation of human hepatocellular carcinoma cell line SMMC-7721. Apoptosis was detected by Annexin V-FITC / PI double-labeled flow cytometry. The levels of IL-1β, TNF-α in liquid samples were determined by Western blot. The expression of C / EBPβ in human hepatoma cells was detected by Western blot. Results Sinomenine hydrochloride had a significant inhibitory effect on the growth of human hepatocellular carcinoma cells in a dose-and time-dependent manner. The differences were statistically significant (P <0.05) The apoptotic rates of hepatoma cells in each group were (8.31 ± 0.81)%, (12.62 ± 1.13)%, (27.43 ± 0.97)% and (51.74 ± 0.85)%, respectively )%, The difference between the experimental group and the control group was statistically significant (P <0.05); also found that, compared with the negative control group, sinomenine in the role of SMMC-7721 cell supernatant IL-1β, TNF- α levels were significantly decreased, while the expression of C / EBPβ significantly increased (P <0.05). Conclusion Sinomenine hydrochloride can inhibit cell proliferation and promote apoptosis in human hepatocellular carcinoma cell line SMMC-7721, which may be related to the inhibition of the expression of transcription factors C / EBPβ and inflammatory cytokines such as TNF-α and IL-1β, The role of micro-environment.