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Objective:To observe the effects of AFP gene silencing by siRNA on the Survivin mRNA of hepatocellular carcino-ma cell line HepG2. Methods:AFP gene expression was downregulated in HepG2 cell by RNAi, and the AFP content in the superna-tant was detected by ELISA. Survivin mRNA level was tested by RT-PCR. MTT was applied to evaluate cell proliferation. Flow cytom-etry was employed to observe cell apoptosis. Results:At 48 h after transfection, AFP expression was almost completely inhibited, cell proliferation activity was decreased by 43.1%, cell apoptosis rate was increased by 24.3%, and the Survivin mRNA expression was re-duced to 22.0%in the experimental group. No evident changes were observed in negative control and blank groups. Conclusion:AFP gene silenced by RNAi induces growth inhibition and apoptosis promotion of hepatocellular carcinoma cell line HepG2. This gene may be associated with the suppression of Survivin mRNA.
RESUMO
<p><b>OBJECTIVE</b>To investigate the effect of total flavonoids of Scutellaria baicalensis Georgi (TFSB) on exogenous expression of influenza A virus nucleoprotein (NP) in HeLa cells.</p><p><b>METHODS</b>HeLa cells were transiently transfected with the empty vector pcDNA3.1(+) or pcDNA3.1(+)/NP vector harboring influenza A virus NP. The pcDNA3.1(+)/NP-transfected cells were treated with TSFB and the expression of influenza A virus NP in the cell supernatant was measured using colloidal gold immunochromatography 48 h after the transfection; fluorescent quantitative RT-PCR was performed to measure the starting copy number of NP gene.</p><p><b>RESULTS</b>The cells transfected with pcDNA3.1 (+)/NP with and without TFSB treatment were positive for NP expression. Fluorescent quantitative RT-PCR showed that the starting copy number of NP gene in pcDNA3.1(+)/NP-transfected cells was (8.90±2.53)×10⁶ copies/µl, showing no significant difference from that of (6.15±1.49)×10⁶ copies/µl in pcDNA3.1(+)/NP-transfected cells with subsequent TFSB treatment (P>0.05).</p><p><b>CONCLUSION</b>TFSB treatment does not obviously affect exogenous influenza A virus NP gene expression or its protein synthesis in HeLa cells.</p>