Screening of proliferation related lncRNAs in leukemia cell lines by lentivirus shRNA library combined with second-generation sequencing.

Qiuyi MA; Deyang Shi; Bichen Wang; Mutian Cao; Haoyuan LI; Weiping Yuan; Yajing Chu

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Screening of proliferation related lncRNAs in leukemia cell lines by lentivirus shRNA library combined with second-generation sequencing.

Author: Qiuyi MA ¹ ; Deyang SHI ¹ ; Bichen WANG ¹ ; Mutian CAO ¹ ; Haoyuan LI ¹ ; Weiping YUAN ¹ ; Yajing CHU ¹
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1. State Key Laboratory of Experimental Hematology, National Clinical Research Center for Blood Diseases, Haihe Laboratory of Cell Ecosystem, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin 300020, China.
Publication Type:Journal Article
Keywords: high-throughput sequencing; leukemia cell lines; lncRNA C20orf204-203; shRNA library
MeSH: Caspase 3; Cell Line, Tumor; Cell Proliferation/genetics*; Humans; Lentivirus/genetics*; Leukemia/genetics*; Proto-Oncogene Proteins c-bcl-2; RNA, Long Noncoding/metabolism*; RNA, Messenger; RNA, Small Interfering/genetics*
From: Chinese Journal of Biotechnology 2022;38(9):3406-3418
CountryChina
Language:Chinese
Abstract: Long non-coding RNA (lncRNA) has become an important regulator of many cellular processes, including cell proliferation. Although studies have shown that a variety of lncRNAs play an important role in the occurrence and development of hematopoietic malignancies, a more comprehensive and unbiased method to study the function of lncRNAs in leukemia cell lines is lacking. Here, we used short hairpin RNA (shRNA) library combined with high-throughput sequencing to screen lncRNAs that may affect the proliferation of leukemia cell lines, and identified lncRNA C20orf204-203 among 74 candidate lncRNAs in this study. Further experiments showed that C20orf204-203 was localized in the cytoplasm in both K562 and THP-1 cell lines. C20orf204-203 knockdown decreased the proliferation of K562 and THP-1 cell lines accompanied with the increased proportion of early apoptotic cells. We observed the increased mRNA level of BAD gene while decreased protein level of TP53 and BCL2. The expression of Caspase 3 decreased and Caspase 3-cleaved protein increased in THP-1 cell line. However, their changes were inconsistent in the two cell lines. Our experimental results showed that knockdown of lncRNA C20orf204-203 in leukemia cell lines affected cell proliferation although the mechanism of action in different cell lines may differ. Importantly, our research demonstrated the feasibility of using shRNA library combined with high-throughput sequencing to study the role of lncRNA in leukemia cell lines on a large scale.