Mechanism Of PI3K/AKT Signaling Pathway In Cell Transformation Process Of Theileria Annulata

Theileria annulata is an intracellular parasite of the parapodium family,which can be parasitized in infected bovine monocytes,lymphocytes,dendritic cells and erythrocytes.Bovine monocytes parasitized by Theileria annulata show tumor growth characteristics,that is,transformation characteristics when cultured in vitro,but these transformed cells will enter the apoptosis process of normal cells after being treated with buparvaquone(BW720).In order to reveal the mechanism of host cell transformation mediated by Theileria annulata,by analyzing the transcriptome data of host cells infected with Theileria annulata,it was found that genes related to PI3K/AKT/mTOR signaling pathway were enriched in the transformed cells,and that the upstream molecule PI3KR3 of this pathway was highly expressed in the transformed cells.Therefore,this paper makes an in-depth study on PI3KR3 and PI3K/AKT/mTOR signal pathways to explore the mechanism of PI3KR3 and PI3K/AKT/mTOR signaling pathways in the transformation process of Theileria annulata.The research results of this paper are as follows:1.Transcriptional analysis of host cells infected with Theileria annulataThrough transcriptional sequencing analysis of mononuclear cells before and after infection,some signal pathways related to tumor,such as PI3K,AKT,Rapl and Ras,were screened.Among them,the PI3K/AKT/mTOR signal pathway is most obviously enriched in the transformed cells of Theileria annulata,and PI3KR3,AKT and mTOR genes which are related to this signal pathway and play an important role in cell proliferation are screened.2.Effects of drugs(BW720)and inhibitors(LY294002)on PI3KR3After BW720 treatment,it was found that obvious apoptosis was found in the transformed cells,and the transcription and protein levels of PI3KR3 decreased,indicating that BW720 had an inhibitory effect on the proliferation of transformed cells.At the same time,LY294002,a specific inhibitor of PI3KR3,was used to inhibit the expression of PI3KR3 in transformed cells.It was found that the proliferation rate of transformed cells slowed down with the decrease of PI3KR3 expression,and apoptosis appeared,indicating that PI3KR3 can regulate the proliferation of transformed cells.3.Effects of overexpression and interference of PI3KR3 on transformed cells of Theileria annulataPI3KR3 was overexpressed in transformed cells of T.annulus by lentivirus packaging technique.It was found that overexpressed PI3KR3 could regulate the expression of its downstream gene mTOR through AKT,thus promoting the proliferation of infected cells.Progress interferes with the expression of PI3KR3 in transformed cells by siRNA technology.The results showed that after interfering with PI3KR3,the transcriptional and protein levels of downstream genes AKT and mTOR decreased,and the cells proliferated slowly and apoptosis.The results showed that PI3KR3 could regulate the proliferation of transformed cells by AKT and mTOR genes.In summary,PI3KR3 plays a key regulatory role in the transformation of host cells,which lays a foundation for further understanding the relationship between the infected cells and tumors and studying the molecular mechanism of the pathogenesis of Theileria annulata.