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利用大规模平行测序技术鉴定 Hela 细胞中的 piRNAs。

Identification of piRNAs in Hela cells by massive parallel sequencing.

机构信息

Department of Medical Genetics, West China Hospital, Sichuan University, Chengdu, China.

出版信息

BMB Rep. 2010 Sep;43(9):635-41. doi: 10.5483/BMBRep.2010.43.9.635.

Abstract

Piwi proteins and Piwi-interacting RNAs (piRNAs) have been implicated in transposon control in germline from Drosophila to mammals. To examine the profile of small RNA expression in human cancer cells and explore difference in small RNA transcriptome, small RNA libraries prepared from wildtype, HILI overexpressed and HILI knockdowned Hela cells were sequenced using Solexa technology. piRNAs and other repeat- associated small RNAs were observed in Hela cells. By using in situ hybridization, piR-49322 was localized in the nucleolus and around the periphery of nuclear membrane in Hela cells. Following the overexpression of HILI, the retrotransposon elements LINE1 was significantly repressed, while LINE1-associated small RNAs decreased in abundance. The present study demonstrated that HILI along with piRNAs plays a role in LINE1 suppression in Hela cancer cell line.

摘要

Piwi 蛋白和 Piwi 相互作用的 RNA(piRNAs)已被证实参与了从果蝇到哺乳动物的生殖细胞中转座子的控制。为了研究人类癌细胞中小 RNA 表达的特征,并探讨小 RNA 转录组的差异,使用 Solexa 技术对野生型、HILI 过表达和 HILI 敲低的 Hela 细胞的小 RNA 文库进行了测序。在 Hela 细胞中观察到 piRNAs 和其他重复相关的小 RNA。通过原位杂交,piR-49322 在 Hela 细胞的核仁中以及核膜周围定位于核仁中。HILI 过表达后,逆转录转座子元件 LINE1 显著受到抑制,而与 LINE1 相关的小 RNA 丰度降低。本研究表明,HILI 与 piRNAs 一起在 Hela 癌细胞系中发挥抑制 LINE1 的作用。

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