The Effects Of Gene Silence ATF5 Combined With Ganciclovir On U87 Cell Proliferation Of HCMV Positive And Virus Replication

Glioma is the most common primary central nervous system tumor.Despite aggressive treatment,but the prognosis is still poor.In glioma environment,the expression of human cytomegalovirus genes and transcriptions increases with malignancy of tumor.Activating transcription factor 5 is a marker transcription factor of tumor and is highly expressed in glioma.In this paper,we explored the effects of gene silence ATF5 combined with ganciclovir on U87 cell proliferation of HCMV positive and virus replication under the environment of glioma.We screened appropriate concentration of GCV,and then we constructed the LV-ATF5-RNAi U87 cells,a stable cell lines of ATF5 silencing,by lentivirus-mediated small interfering RNA targeting ATF5.LV-ATF5-RNAi U87 cells were HCMV(low multiplicity of infection)infected,cultivated in medium containing GCV.Under the condition of gene silence ATF5 combining with GCV,cell proliferation was detected by CCK-8 assay,the expression level of HCMV gene and protein(IE,UL44 and UL99)was analyzed by Real-time PCR and Western-blot,respectively,and the level of viral progeny production was tested by Plaque-forming assay.The results showed that,the stable cell line of LV-ATF5-RNAi U87 with ATF5 silencing had constructed successfully.Gene silence ATF5 combining with GCV showed the better inhibition on HCMV positive glioma.Concordant results showed gene silence ATF5 combining with GCV displayed the most potent inhibition effects in HCMV mRNA expression,protein expression and viral progeny production,both stronger than GCV or ATF5 gene silencing action alone.The results suggest that gene silence ATF5 combining with GCV can be suppress HCMV viral production and malignant proliferation of glioma.So,gene silence ATF5 combining with GCV may be a new way for clinical treatment of glioma.