Effects Of Interferon-α On Gene Expression Profile In Human Nerve Cell Line SH-SY5Y Cells

BackgroundInterferon-α (IFN-α) is one of the classic innate immune cytokines widely used in antiviral (HCV, HBV) and antitumor (melanoma, breast carcinoma) therapies. However, IFN-α often causes individuals to experience significant psychiatric side effects, such as depression, fatigue, insomnia, anxiety, and cognitive disturbances if the treatment is continued for a long time. It is estimated that approximately 30%-70% of hepatitis C virus-infected patients treated with IFN-a experience different degrees of depression. Such side effects involving depression can lead to interruption of IFN-a treatment in patients6. It was very necessary to develop effective methods to monitor or predict the onset of depression in the paints received IFN-a treatment and thus avoid the development of depression. we suspected to develop the methods through some genetic biomarker (susceptibility gene) to predict the risk of depression.ObjectiveLooking for an objective and accurate biological sign (susceptive gene) which can predict the risk of depressionMethods1.Giving medicine:the final concentration of INF-ais 3000IU/M1. Putting it in the SH-sy5y cell culture medium and culturing 16h, at the same time, setting the control group which is putting PBS in the medium.2. mRNA extraction:Using the method of Trizol extract RNA from the cell.3. Reverse transcription for cDNA:Making RNA which is extracted from SH-SY5Y cell reverse transcription for cDNA.4. Gene Chip detection:Separating detect the expression of 5-HT and Dopamine metabolism related gene、inflammation related gene、Neural growth related gene Classical depression related gene. Repeated three times.5. Statistical analysis:We use spssl9.0 statistical software analyzing the result through T-test. P<0.05 signify the significant difference. The screening condition of susceptibility genes of depression is that the ratio of spotting group to control group is >2.0 or <0.5 (P<0.05).Results1. The signal of All the entire gene expression profile chip is even and clear which is not marginal effect,、scratches and other impact signal value. The results of positive anchor point and negative control area are obvious.2. The gene expression profile chip clustering figure shows that the experiment sample of each group cluster together through three times repeated.The result proves that the experimental repeatability.3.INF-a can improve the genetic expression level of TPH2、CASP1、INDO、IL7、 IL12A.IL15 RNASH-SY5Y in SH-SY5Y cell.ConclusionINF-a can improve the genetic expression level of TPH2、CASP1、INDO、IL7、IL12A、IL15 RNASH-SY5Y in SH-SY5Y cell.