Liu Zhaoping, Wang Yanyan, Xu Zhenru, Yuan Shunling, Ou Yanglin, Luo Zeyu, Wen Feng, Liu Jing, Zhang Ji
Department of Clinical Laboratory, Shenzhen Traditional Chinese Medicine Hospital, Shenzhen, Guangdong, China.
Department of Clinical Laboratory, The First Affiliated Hospital, University of South China, Hengyang, Hunan, China.
PeerJ. 2021 May 25;9:e11429. doi: 10.7717/peerj.11429. eCollection 2021.
Drug resistance is the main obstacle in the treatment of leukemia. As a member of the competitive endogenous RNA (ceRNA) mechanism, underlying roles of lncRNA are rarely reported in drug-resistant leukemia cells.
The gene expression profiles of lncRNAs and mRNAs in doxorubicin-resistant K562/ADR and sensitive K562 cells were established by RNA sequencing (RNA-seq). Expression of differentially expressed lncRNAs (DElncRNAs) and DEmRNAs was validated by qRT-PCR. The potential biological functions of DElncRNAs targets were identified by GO and KEGG pathway enrichment analyses, and the lncRNA-miRNA-mRNA ceRNA network was further constructed. K562/ADR cells were transfected with CCDC26 and LINC01515 siRNAs to detect the mRNA levels of GLRX5 and DICER1, respectively. The cell survival rate after transfection was detected by CCK-8 assay.
The ceRNA network was composed of 409 lncRNA-miRNA pairs and 306 miRNA-mRNA pairs based on 67 DElncRNAs, 58 DEmiRNAs and 192 DEmRNAs. Knockdown of CCDC26 and LINC01515 increased the sensitivity of K562/ADR cells to doxorubicin and significantly reduced the half-maximal inhibitory concentration (IC) of doxorubicin. Furthermore, knockdown of GLRX5 and DICER1 increased the sensitivity of K562/ADR cells to doxorubicin and significantly reduced the IC of doxorubicin.
The ceRNA regulatory networks may play important roles in drug resistance of leukemia cells. CCDC26/miR-140-5p/GLRX5 and LINC01515/miR-425-5p/DICER1 may be potential targets for drug resistance in K562/ADR cells. This study provides a promising strategy to overcome drug resistance and deepens the understanding of the ceRNA regulatory mechanism related to drug resistance in CML cells.
耐药性是白血病治疗的主要障碍。作为竞争性内源性RNA（ceRNA）机制的成员，长链非编码RNA（lncRNA）在耐药白血病细胞中的潜在作用鲜有报道。
通过RNA测序（RNA-seq）建立阿霉素耐药K562/ADR细胞和敏感K562细胞中lncRNA和mRNA的基因表达谱。通过qRT-PCR验证差异表达lncRNA（DElncRNA）和差异表达mRNA（DEmRNA）的表达。通过GO和KEGG通路富集分析确定DElncRNA靶标的潜在生物学功能，并进一步构建lncRNA-miRNA-mRNA ceRNA网络。用CCDC26和LINC01515 siRNA转染K562/ADR细胞，分别检测GLRX5和DICER1的mRNA水平。用CCK-8法检测转染后的细胞存活率。
基于67个DElncRNA、58个DEmiRNA和192个DEmRNA，ceRNA网络由409个lncRNA-miRNA对和306个miRNA-mRNA对组成。敲低CCDC26和LINC01515可增加K562/ADR细胞对阿霉素的敏感性，并显著降低阿霉素的半数最大抑制浓度（IC）。此外，敲低GLRX5和DICER1可增加K562/ADR细胞对阿霉素的敏感性，并显著降低阿霉素的IC。
ceRNA调控网络可能在白血病细胞耐药中起重要作用。CCDC26/miR-140-5p/GLRX5和LINC01515/miR-4俯5-5p/DICER1可能是K562/ADR细胞耐药的潜在靶点。本研究为克服耐药性提供了一种有前景的策略，并加深了对慢性粒细胞白血病细胞中与耐药相关的ceRNA调控机制的理解。
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