伊马替尼耐药K562/G的microRNA表达谱分析及与FOXO3/Bcl-6信号通路关系的研究  

The Relationship between MicroRNA Expression Profiling in Imatinib-Resistant Cell Line K562/G and Potential Mechanism through FOXO3/Bcl-6 Signaling Pathway

作　　者：沈晶[1] 王欢[1] 王晶石[1] 魏娜[1] 王昭[1] SHEN Jing;WANG Huan;WANG Jing-Shi;WEINa;WANG Zhao(Department of Hematology,Beijing Friendship Hospital Affiliated to Capital Medical University,Beijing 100050,China)

机构地区：[1]首都医科大学附属北京友谊医院血液内科,北京100050

出　　处：《中国实验血液学杂志》2022年第1期107-112,共6页Journal of Experimental Hematology

基　　金：国家自然科学基金资助项目(81200393)。

摘　　要：目的:探讨CML耐药细胞株K562/G耐药相关的FOXO3/Bcl-6信号通路及相关microRNA(miRNA)机制。方法:采用MTT法检测伊马替尼对K562/G的耐药性;采用Western blot法检测耐药株和敏感株细胞中FOXO3和Bcl-6蛋白的表达情况,实时荧光定量PCR检测FOXO3及Bcl-6 mRNA的表达。运用miRNA芯片技术筛查K562与K562/G细胞之间差异表达的miRNA,进一步筛选FOXO3/Bcl-6信号通路的靶向miRNA。结果:K562/G较K562细胞株的FOXO3及Bcl-6蛋白表达均明显升高(P<0.01);Bcl-6 mRNA的表达水平无明显差异,而K562/G细胞中FOXO3 mRNA表达明显升高(P<0.05)。miRNA芯片结果显示,K562/G与K562细胞之间有109条miRNA存在显著的差异,其中上调miRNA为81个,下调miRNA有28个。经生物信息学反向预测,其中miR-6718-5p、miR-5195-5p、miR-4711-3p、miR-4763-5p、miR-4664-5p和miR-3176与该信号通路相关。结论:伊马替尼耐药与FOXO3/Bcl-6信号通路相关,并且K562/G与K562细胞存在miRNA表达显著差异。Objective:To investigate the drug resistant related FOXO3/Bcl-6 signaling pathway in K562/G cell line and its related microRNA(miRNA) mechanisms.Methods:The drug resistance potency of imatinib on K562/G was detected by MTT assay.The expression of FOXO3 and Bcl-6 proteins in K562 and K562/G cells was detected by Western blot.Real-time PCR(RT-PCR) was used to detect the expression of FOXO3 and Bcl-6 mRNA.The miRNA expression profiling in K562 and K562/G cells was analyzed by micro array technique,and the miRNA targeted to FOXO/Bcl-6 signaling pathway was identified.Results:The expression of FOXO3 and Bcl-6 protein was significantly increased in K562/G cells as compared with that in K562 cells(P <0.01),the expression level of Bcl-6 mRNA showed no increase in K562/G cells.However,FOXO3 mRNA was up-regulated in K562/G cells(P <0.05).MiRNA microarray results showed that 109 miRNAs were expressed differentially in K562 and K562/G cells.The expression of81 miRNA s were up regulated while 28 miRNAs were down-regulated.Through reverse prediction by bio informatics,miR-6718-5 p,miR-5195-5 p,miR-4711-3 p,miR-4763-5 p,miR-4664-5 p and miR-3176 were related to FOXO/Bcl-6 signaling pathway.Conclusion:The FOXO3/Bcl-6 signaling pathway contributes to imatinib resistance in K562/G cell line,and the miRNA expression profiles showed significant differences between K562/G and K562 cells.

关 键 词：慢性髓系白血病 耐药 miRNA FOXO3 BCL-6 

分 类 号：R733.72[医药卫生—肿瘤]