基于转录组数据分析环形泰勒虫对诱导宿主细胞转化相关基因的影响

doi:10.11843/j.issn.0366-6964.2022.12.025

Abstract

Abstract: This study was conducted to study the changes of transcriptome in Theileria annulata infected bovine monocytes (TaNM Ⅰ) before and after treatment with Bupavaquone (BW720) for laying a foundation for further study on the molecular mechanism of host cell transformation induced by T. annulata. Bovine TaNM Ⅰ cells infected by T. annulata were used as experimental materials, control group (DMSO) and experimental group (BW720) were set up. High-throughput sequencing was carried out by Illumina HiSeq4000 sequencing platform. The original reading segment (Raw reads) was compared with GenBank and Rfam database, and the clean value (Clean reads) was obtained by quality control. The differentially expressed genes in DMSO treatment group and BW720 treatment group were screened by Venn analysis software, and functional annotation (GO) and signal pathway (KEGG) analysis were performed. Ten genes were randomly selected and the expression of genes in cells under different treatment conditions was detected by fluorescence quantitative PCR (qPCR). After verifying the accuracy of the sequencing results, the differentially expressed genes were selected and their roles in apoptosis and proliferation of TaNM Ⅰ cells were analyzed. Results showed that 6 854 019 704 and 6 627 265 854 Raw reads were obtained in BW720 treatment group and DMSO treatment group, and 22 925 606 and 22 171 427 Clean read were obtained after filtration and quality control, respectively. A total of 4 054 genes were expressed in BW720 group and DMSO group, of which 2 146 genes were significantly up-regulated and 1 908 genes were significantly down-regulated. Three hundred and sixty-seven differentially expressed genes were further screened, of which 196 were significantly up-regulated and 171 were significantly down-regulated. At the same time, the expression trend verified by qPCR was consistent with that of transcriptome sequencing, indicating that the sequencing results were reliable. Then GO functional annotation was performed on the differentially expressed genes, and 20 biological processes, 15 cell components and 11 molecular functional items related to cell proliferation were screened. KEGG enrichment analysis showed that some signaling pathways related to T. annulata transformation cells were screened out in Top20 signaling pathway, such as cancer signaling pathway, PI3K-Akt signaling pathway, MAPK signaling pathway, etc. The progress analysis of these signaling pathways found that PI3KR3, FOXO1, IL23A, FZD3, AKT, MMP9 and other genes have also been reported in the study of transformation cells of T. annulata. This study shows that DAPK1,FZD3,FOXO3 and PI3KR3 may be the key candidate genes affecting the infinite proliferation of infected cells in TaNM Ⅰ cells. It lays a foundation for the follow-up study of the mechanism of infinite proliferation of TaNM Ⅰ cells.

Key words: T. annulata, bovine monocyte, transcriptome, signal pathways

CLC Number:

S852.723

ZHAO Baocai, ZHAO Shuaiyang, GUAN Guiquan, LUO Jianxun, CAO Tianxing, ZHANG Zhigang, SHI Miao, LIU Junlong, ZHAO Hongxi. Transcriptome-based Analysis of the Effects of Theileria annulata on Host Cell Transformation-related Genes[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(12): 4398-4409.

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Transcriptome-based Analysis of the Effects of Theileria annulata on Host Cell Transformation-related Genes

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