Effect Of MicroRNA-141-3p On Cell Proliferation And Apoptosis Of Glioma Via Targeting ATF5

Glioma is the most common malignant primary brain tumor which arises from the central nervous system.Glioblastoma is considered as the most aggressive malignant glioma.MicroRNAs(miRNAs)are a kind of endogenous,non-coding,small(19-25 nucleotides)RNAs,which bind to the 3'-UTR of target gene mRNAs to negatively regulate gene expression on a post-transcriptional level.It participates in physiological processes such as individual development,apoptosis,proliferation,and differentiation.Loss or abnormal expression of these miRNAs may be closely related to disease,including tumors.However,the specific mechanism of the occurrence and development of tumors by miRNAs is unknown.Our team has detected that HCMV infection is associated with proliferation and apoptosis of glioma cells.And some data showed that the expression of miR-141-3p after HCMV infection was significantly reduced.Therefore,we hypothesized that HCMV affects the malignant degree of glioma cells by regulating the expression of miRNA in glioma cells and controlling the key proteins that modulated cell proliferation and apoptosis in gliomas.Bioinformatic analysis showed that ATF5 was a potential target of miR-141-3p.To further improve the reliability of the prediction,we used the BiBiServe RNA hybrid algorithm to predict the minimum free energy of binding of miR-141 ATF5 3'-UTR.The predicted results show that the free energy of binding of miR-141 to the ATF5 3'-UTR region is-74.0 kcal/mol,and there are seven nucleotides in the seed region that are fully complementary.Luciferase reporter assay verified that miR-141-3p specifically targeted the ATF5 3'-UTR in glioma cells.The expression of miR-141-3p and ATF5 in glioma tissue samples and cells were detected by real-time PCR,and the effect of miR-141-3p on ATF5 mRNA and protein expression was examined.The results show that miR-141-3p can reduce ATF5 mRNA and protein expression.By CCK8 assay and Annexin V-PE / 7-AAD assay,we further found that miR-141-3p negatively regulates ATF5,then inhibiting cell proliferation and promoting apoptosis.Our results confirm that the seed sequence of miR-141-3p is extremely important for its binding to the ATF5 3'-UTR to form a secondary structure,and this binding inhibits the further translation of the ATF5 mRNA.It's not only provides a new possible target for targeting glioma therapy,but also provides a new theoretical and experimental basis for clarifying the development mechanism of glioma.At the same time,it is predicted that HCMV infection is likely to use the regulation of a variety of expression of MicroRNA to enhance the continuous proliferation of tumor cells,in order to create conditions for continuous infection.This will be the focus of our research team's next research.