目的 利用RNA-seq技术分析刚地弓形虫入侵人包皮成纤维（HFF）细胞前后转录组的差异。方法 将纯化的刚地弓形虫RH株速殖子与HFF细胞按3 : 1的比例共培养,37 ℃、5% CO₂培养24 h后收集HFF细胞和弓形虫共培养物（感染组）,以纯化弓形虫RH株速殖子作为对照组。提取感染组和对照组总RNA,纯化mRNA并构建转录组文库,利用BGISEQ-500平台进行高通量测序,测序获得的质控数据与NCBI数据库中的弓形虫基因组（GCF_000006565.2_TGA4_ncbi）进行比对,对感染组与对照组相比差异倍数 ≥ 2.0且P < 0.05的差异表达基因进行功能注释、基因本体（GO）功能富集分类、富集和京都基因与基因组百科全书(KEGG)通路分析以及蛋白相互作用网络分析。结果 共测序获得5 888个基因,差异基因986个,其中上调基因533个,下调基因453个。GO分析结果显示,富集的GO条目分别为膜、膜的组成部分、膜的固有成分、膜相关组分等。KEGG富集通路主要为胃癌、脂肪酸生物合成、钙离子信号等通路。蛋白相互作用网络分析结果显示,相互作用蛋白数最多的6个蛋白分别是TGME49_282200、TGME49_305980（PDHE3I）、TGME49_316310（SOD）、TGME49_310440（MORN1）、TGME49_237110和TGME49_222020（PGKII）。结论 弓形虫RH株速殖子入侵HFF细胞前后转录组存在差异,差异表达基因KEGG富集通路主要为胃癌、脂肪酸生物合成、钙离子信号等通路,筛选出6个相互作用蛋白数量最多的蛋白基因。

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.