参芪扶正注射液联合IFN-α对肝癌细胞STAT1基因表达的影响

摘要

Objective:To study the effect of Shenqi Fuzheng injection combined with interferon-α (IFN-α) on the expression of STAT1 gene in hepatocellular carcinoma cells,and to elucidate its mechanism of IFN-α synergistic effect.Methords:The effect of IFN-α injection alone or combined with Shenqi Fuzheng injection on the proliferation of MHCC97-L cells was detected by MTT assay.IFN-α was detected by Real-time PCR and Western-blot respectively.The expression of STAT1 mRNA and protein in the experimental group,the negative control group (NaCl) and the blank control group were determined,and the effect of Shenqi Fuzheng injection on the transcription and expression of STAT1 was determined.The expression of STAT1 in lentiviral vector MTT assay was used to detect the effect of IFN-α alone or in combination with Shenqi Fuzheng injection on the STAT1 gene "silencing" cells.The expression of STAT1 was detected by RT-PCR and Western-Blot in MHCC97L cells.Strain,on cell proliferation.Results:Compared with IFN-α alone,Shenqi Fuzheng injection combined with IFN-α enhanced the inhibitory effect of IFN-α on MHCC97-L (P＜0.01) and up-regulated the expression of STAT1mRNA and protein (P＜0.05).Successfully constructed the STAT1 gene "silent" in the MHCC97-L cell line.There was no significant difference in the inhibition rate of MHCC97-L between Shenqi Fuzheng injection and IFN-α (P＞0.05).Conclusion Shenqi Fuzheng Injection can up-regulate the expression of STAT1 in human hepatocellular carcinoma cell line MHCC97-L,so as to increase the effect of IFN-α.%目的:研究参芪扶正注射液与干扰素α (IFN-α)联用时对肝癌细胞STAT1基因表达的影响,探讨其对IFN-α协同增效的作用机制.方法:运用MTT法检测IFN-α注射液单独或与参芪扶正注射液联合对MHCC97-L细胞增殖的影响;运用Real-time PCR和Western-blot分别检测IFN-α单独或与参芪扶正注射液联用对STAT1 RNA和蛋白质表达的影响;构建STAT1干扰慢病毒载体并实现其在MHCC97-L中表达,通过RT-PCR、Western-Blot验证其干扰STAT1表达的有效性;MTT法检测IFN-α单独或联合参芪扶正注射液对STAT1基因“沉默”细胞株增殖的影响.结果:与单独使用IFN-α相比,参芪扶正注射液与IFN-α联用可以增强IFN-α对人肝癌MHCC97-L的抑制作用(P＜0.01),并上调STAT1 mRNA(P＜0.05)和蛋白质的表达(P＜0.05);成功构建STAT1基因“沉默”的MHCC97-L细胞株;参芪扶正注射液协同IFN-α对STAT1基因“沉默”MHCC97-L的抑制率与单独使用IFN-α差异无统计学意义(P＞0.05).结论:参芪扶正注射液可上调入肝癌细胞MHCC97-L中STAT1的表达,从而提高IFN-α的疗效.