Over-expression of hsa-miR-221 in JSC1 cells and identification of miRNA targets expressed in a panel of B-cell lines. Homo sapiens

1) To identify changes in gene expression upon over-expression of hsa-miR-221 in JSC1 cells. 2) To identify human miRNA targets expressed in a panel of B-cell lines. Gammaherpesvirus and host cell microRNAs (miRNAs) together modulate gene expression in normal and malignant cells. Using microRNA microarrays, we determined the expression of mature viral and host cellular miRNAs in a series of B cell tumours that include Kaposi’s Sarcoma-associated herpesvirus (KSHV) infected Primary Effusion Lymphoma (PEL) and Epstein-Barr virus (EBV) infected Burkitt’s lymphoma (BL) cell lines. We show that 35 host miRNAs were constitutively expressed in all the B cell lymphomas and differences in viral miRNA expression were evident between herpesvirus positive tumour types. Furthermore, we show that in PEL, miR-221 and miR-222 expression is defective due to a lack of transcript expression rather than mutation in the miRNA encoding loci. Absence of miR-221 and miR-222 resulted in the enhanced expression of the known target gene p27 (CDKN1B) and reintroduction of miR221 in PEL reduces p27 protein expression. Overall design: The dataset comes in two parts. 1) 6 JSC1 samples: 2 with miR-221 lentiviral vector, 2 without and 2 with K5 short hairpin RNA as control. 2) A panel of 14 B-cell line samples. Each of the 20 samples was Cy5 labelled and run with Cy3-labelled Stratagene reference RNA.

本数据集包含两项研究目标：1）鉴定JSC1细胞中过表达hsa-miR-221后的基因表达变化；2）鉴定在一组B细胞系中表达的人类微小核糖核酸（microRNA, miRNA）靶标。
伽马疱疹病毒与宿主细胞miRNA共同调控正常及恶性细胞内的基因表达。我们通过微小核糖核酸微阵列（microRNA microarray），分析了一系列B细胞肿瘤中成熟病毒及宿主细胞miRNA的表达情况，这些肿瘤包括卡波西肉瘤相关疱疹病毒（Kaposi’s Sarcoma-associated herpesvirus, KSHV）感染的原发性渗出性淋巴瘤（Primary Effusion Lymphoma, PEL），以及爱泼斯坦-巴尔病毒（Epstein-Barr virus, EBV）感染的伯基特淋巴瘤（Burkitt’s lymphoma, BL）细胞系。研究发现，35种宿主miRNA在所有B细胞淋巴瘤中均呈组成型表达，且疱疹病毒阳性的不同肿瘤类型之间，病毒miRNA的表达存在显著差异。此外，我们证实PEL细胞中miR-221与miR-222的表达存在缺陷，其原因并非miRNA编码位点发生突变，而是缺乏转录本的表达。miR-221与miR-222的缺失会导致已知靶基因p27（CDKN1B）的表达上调，而在PEL细胞中重新导入miR-221则可降低p27蛋白的表达水平。

整体实验设计：本数据集分为两部分。1）6份JSC1细胞样本：2份转染miR-221慢病毒载体、2份未转染（空白对照），以及2份转染K5短发夹RNA作为对照。2）一组共14份B细胞系样本。所有20份样本均经Cy5荧光标记，并与Cy3标记的Stratagene参考RNA进行杂交实验。