Expression profiles of mRNA and lncRNA in HCT-8 cells infected with Cryptosporidium parvum IId subtype. Expression profiles of mRNA and lncRNA in HCT-8 cells infected with Cryptosporidium parvum IId subtype

Cryptosporidium parvum is one of the most important opportunistic enteric parasites, causing severe diarrhea in immunocomprised human and animals. However, few effective control agents were available for this parasite. Long non-coding RNA (lncRNA) was discovered to play key roles in many diseases through regulating the gene expression. Here, using microarray assay, we investigated the expression profiles of mRNA and lncRNA in HCT-8 cells after the infection of C. parvum IId subtype, the prevalent subtype of China. A total of 821 lncRNAs and 1,349 mRNAs were dysregulated expressed in HCT-8 cells at 24 h post infection (pi) of C. parvum IId subtype. Of them, all five types of lncRNAs were identified, including 302 of sense, 280 of antisense, 312 of intergenic, 44 of divergent, and 33 of intronic lncRNAs. Ten lncRNAs and ten mRNAs randomly selected were successfully confirmed the microarray results. The co-expression and target prediction analysis indicated 27 of mRNAs cis-regulated by 29 lncRNAs and 109 of coding genes trans-regulated by 114 lncRNAs. These predicted targets were enriched in pathways involved in the interaction between host and C. parvum, eg. hedgehog signaling pathway, Wnt signaling pathway and tight junction, suggesting that these aberrantly lncRNAs would play important regulating roles during infection of C. parvum IId subtype. Overall design: we collected the HCT-8 cells infected with cryptosporidium parvum at 24 hours and analyzed the profiles of mRNAs and lncRNAs.

微小隐孢子虫（Cryptosporidium parvum）是最为重要的机会性肠道寄生虫之一，可引发免疫受损人类与动物出现严重腹泻。然而当前针对该寄生虫的有效防控制剂极为匮乏。长链非编码RNA（long non-coding RNA，lncRNA）已被证实可通过调控基因表达，在多种疾病进程中发挥关键调控作用。本研究借助基因芯片检测（microarray assay），分析了中国流行的微小隐孢子虫IId亚型感染HCT-8细胞后，细胞内mRNA与lncRNA的表达谱。在感染微小隐孢子虫IId亚型24小时（感染后，pi）的HCT-8细胞中，共计821个lncRNA与1349个mRNA呈现异常表达。其中共鉴定出5类lncRNA，分别为302个正义链lncRNA、280个反义链lncRNA、312个基因间lncRNA、44个双向lncRNA以及33个内含子lncRNA。研究随机选取10个lncRNA与10个mRNA进行验证，成功确认了基因芯片检测结果的可靠性。共表达与靶基因预测分析显示，29个lncRNA可顺式调控27个mRNA，114个lncRNA可反式调控109个编码基因。上述预测靶基因显著富集于宿主与微小隐孢子虫互作相关的通路，例如刺猬信号通路（hedgehog signaling pathway）、Wnt信号通路（Wnt signaling pathway）以及紧密连接（tight junction），提示这些异常表达的lncRNA在微小隐孢子虫IId亚型感染过程中发挥重要调控作用。实验总体设计：本研究收集感染微小隐孢子虫24小时的HCT-8细胞，对其mRNA与lncRNA的表达谱进行分析。