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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、We s te rn-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人肝癌细胞MHCC97-L中STAT1的表达,从而提高IFN-α的疗效。
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 its mechanism of synergism with IFN-α. Methods: MTT assay was used to detect the effects of IFN-α injection alone or combined with Shenqi Fuzheng injection on the proliferation of MHCC97-L cells. Real-time PCR and Western-blot were used to detect IFN-α alone or with Shenqi Fuzheng injection The effect of STAT1 RNA and protein expression on the STAT1 RNA and protein expression was investigated. STAT1 was constructed and its expression in MHCC97-L was confirmed. The STAT1 expression was verified by RT-PCR and Went te rn-Blot. MTT assay To detect the effect of IFN-α alone or in combination with Shenqi Fuzheng injection on the proliferation of STAT1 gene silenced cell line. Results: Shenqi Fuzheng injection combined with IFN-α could enhance the inhibitory effect of IFN-α on human hepatocarcinoma MHCC97-L (P <0.01) and upregulate STAT1 mRNA (P <0.05) ) And protein (P <0.05). The MHCC97-L cell line with STAT1 gene silencing was successfully constructed. The inhibitory rate of Shenqi Fuzheng injection combined with IFN-α on STAT1 gene silencing MHCC97-L There was no significant difference with IFN-α alone (P> 0.05). Conclusion: Shenqi Fuzheng injection can up-regulate the expression of STAT1 in human hepatocellular carcinoma cell line MHCC97-L, thereby enhancing the therapeutic effect of IFN-α.