Detection Of Epstein-Barr Virus Genes Expressions In Raji Cell Line With Stable Low Expression Of PLK1 Gene

[Objective] Polo-like kinase 1(PLK1)is overexpressed in human tumors.Our previous experiments also found that PLK1 gene was overexpressed in EBV-transformed lymphoblast LCLs.In this experiment,CCK-8 and Hoechst staining were used to observe the proliferation and apoptosis of Raji cells after interfering with the expression of PLK1 gene;The expression of Epstein-Barr virus(EBV)genes in Raji cells with lower expression of PLK1 gene were detected by q RT-PCR and Western blot for further study of the interaction between Epstein-Barr virus and host cells.[Method] Raji cell line with low expression of PLK1 gene was cultured.The green fluorescence of cells was observed by inverted fluorescence microscope.The proliferation and apoptosis of Raji cells were observed by CCK-8 assay and Hoechst staining.The transfection efficiency of Raji cells with low expression of PLK1 gene was verified by q RT-PCR.q RT-PCR and Western Blot were used to detect the expression of EBV genes in Raji cells with lower expression of PLK1 gene.[Results] 1.Raji cell line with low expression of PLK1 gene was cultured.The transfection group(Raji/PLK1 sh RNA and Raji/NC)were observed to have up to 70% of cells carrying green fluorescence by inverted fluorescence microscope;Quantitative real-time PCR detection showed that Raji/PLK1 sh RNA and Raji/NC compared with the blank group(Raji),the expression of PLK1 at the m RNA level was significantly decreased,indicating that the Raji cell line with low expression of PLK1 gene has a good interference effect.2.Interference with PLK1 gene expression could inhibit proliferation and induce cell apoptosis of Raji cellsThe results of CCK-8 showed that the growth rate of Raji/PLK1 sh RNA cells was significantly lower than that in Raji/NC cells and Raji cells(P <0.01),which showed that the interference with PLK1 gene expression was capable of inhibiting the proliferation of Raji cell line.Hoechst staining showed that Raji/NC cells showed weak blue fluorescence & a small amount of red fluorescence,while Raji/PLK1 sh RNA cells showed strong blue fluorescence & a small amount of red fluorescence,Raji/PLK1 sh RNA cells were significantly more than Raji/NC cells and Raji cells,implying that the interference with PLK1 gene expression could induce cell apoptosis of Raji cell line.3.Knockdown the expression of PLK1 gene could affect EBV genes expressionKnockdown the expression of PLK1 gene in Raji cells could up-regulate the expression of EBV genes LMP-1,LMP-2A and EBNA-1.In this study,the expression of PLK1 gene in Raji cells was detected by Quantitative real-time PCR.We found that knocking down the expression of PLK1 in Raji lymphoma cells could up-regulate the expression of LMP-1,LMP-2A and EBNA-1 genes.[Conclusion] The interference of PLK1 gene expression in Raji cells could inhibit the growth of Raji cells,induce apoptosis,and up-regulate the expressions of Epstein-Barr virus LMP-1,LMP-2A and EBNA-1 genes.