Comparative analysis of transcriptomes in Toxoplasma gondii before and after invasion in human foreskin fibroblasts

doi:10.12140/j.issn.1000-7423.2021.04.010

Abstract

Abstract:

Objective To investigate transcriptome changes in Toxoplasma gondii before and after invasion in human foreskin fibroblasts (HFF) by RNA-seq. Methods Tachyzoites of T. gondii RH strain were added to monolayered HFF cells at a parasite-cell infection ratio of 3 : 1 and incubated at 37 ℃ with 5% CO₂ for 24 h. Following the incubation, the cells (infection group) were harvested and RNA extracted, using tachyzoites of RH strain as a blank control (control group). The mRNA was purified for high-throughput sequencing using the BGISEQ-500 platform. The eligible sequence data obtained were aligned with T. gondii genome data (GCF_000006565.2_TGA4_ncbi） in NCBI database. Differentially expressed genes (2.0-fold change with a P-value of < 0.05 in the infection group in comparison with the control group) were selected for Gene Ontology (GO) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment and protein interaction network analysis. Results A total of 5 888 genes were detected, 986 differentially expressed genes were identified, in which 533 upregulated and 453 downregulated. GO analysis showed that the GO items significantly enriched were membrane, integral component of membrane, intrinsic component of membrane, and membrane part. KEGG enrichment pathways mainly involving gastric cancer, fatty acid biosynthesis, and calcium signaling pathway. In protein-protein interaction network analysis, the top six interacting proteins were TGME49_282200, TGME49_305980 (PDHE3I), TGME49_316310 (SOD), TGME49_310440 (MORN1), TGME49_237110 and TGME49_222020 (PGKII). Conclusion There are significant differences in transcriptome in T. gondii before and after invasion in HFF cells. The KEGG enrichment pathways mainly involve gastric cancer, fatty acid biosynthesis, and calcium signaling pathway, and six top genes with regard to the number of interaction proteins are screened.

Key words: Toxoplasma gondii, Transcriptome, Differentially expressed gene

CLC Number:

R382.5

WANG Long-jiang, LI Jin, YIN Kun, XU Chao, LIU Gong-zhen, HUANG Bing-cheng, WEI Qing-kuan, SUN Hui. Comparative analysis of transcriptomes in Toxoplasma gondii before and after invasion in human foreskin fibroblasts[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2021, 39(4): 480-486.

Figures/Tables 4

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Table 1

KEGG pathway analysis of differentially expressed genes in T. gondii before and after infection of HFF cells

序号 No.	通路序号 Pathway ID	代谢通路 Metabolism pathway	差异基因数量 No. of differentially expressed proteins	基因名称 Gene name
1	ko05226	胃癌 Gastric cancer	22	ROP18、GAP50、ALV12、ALV4、GRA15、SUB7、TGME49_208370、TGME49_209050、TGME49_220240、TGME49_220280、TGME49_220640、TGME49_224150、TGME49_226950、TGME49_239020、TGME49_240060、TGME49_254630、TGME49_267450、TGME49_278090、TGME49_278370、TGME49_287960、TGME49_301690、TGME49_320020
2	ko00061	脂肪酸生物合成 Fatty acid biosynthesis	4	ACC1、TGME49_225990、TGME49_231890、TGME49_293590
3	ko04020	钙离子信号通路 Calcium signaling pathway	12	ROP19A、CAM1、PMCAA1、TGME49_209985、TGME49_220420、TGME49_226540、TGME49_297670、TGME49_300360、TGME49_311260、TGME49_318675、TGME49_322000、TGME49_322010
4	ko04024	cAMP信号通路 cAMP signaling pathway	11	IMC2A、CAM1、PMCAA1、TGME49_209050、TGME49_209985、TGME49_220420、TGME49_226540、TGME49_293000、TGME49_294190、TGME49_297670、TGME49_311260
5	ko04144	内吞作用 Endocytosis	13	TGME49_209985、TGME49_212300、TGME49_220460、TGME49_239580、TGME49_242860、TGME49_251170、TGME49_252490、TGME49_262950、TGME49_291800、TGME49_305270、TGME49_313450、TGME49_315210、TGME49_315350
6	ko04657	IL-17信号通路 IL-17 signaling pathway	14	ROP18、IMC6、TGME49_209050、TGME49_212810、TGME49_217540、TGME49_219270、TGME49_220640、TGME49_224150、TGME49_240810、TGME49_250115、TGME49_254630、TGME49_268870、TGME49_271970、TGME49_320150
7	ko04927	皮质醇合成与分泌Cortisol synthesis and secretion	4	TGME49_209985、TGME49_224840、TGME49_293000、TGME49_297670
8	ko04540	间隙连接 Gap junction	5	TUBA1、TGME49_209985、TGME49_212240、TGME49_221620、TGME49_297670
9	ko00300	赖氨酸生物合成 Lysine biosynthesis	2	TGME49_227090、TGME49_234440
10	ko03420	核苷酸切除修复Nucleotide excision repair	6	GTF2H3、PCNA2、TGME49_203170、TGME49_237110、TGME49_254640、TGME49_319860

Table 1

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