LncRNA XIST负调控miR-196b对急性髓系白血病细胞KG1a生物行为的影响  

Effect of Negative Regulation of LncRNA XIST to MiR-196b on the Biological Behavior of Acute Myeloid Leukemia Cells KG1a

作　　者：王筝[1] 马杏凡 万春超 张岚 王静波[1] WANG Zheng;MA Xing-Fan;WAN Chun-Chao;ZHANG Lan;WANG Jing-Bo(Department of Hematology,Bejing Aerospace Center Hospital,Bejing 100049,China;Research Office,Shougang Hospital of Peking University,Beijing 100144,China)

机构地区：[1]航天中心医院血液科,北京100049 [2]北京大学首钢医院科研处,北京100144

出　　处：《中国实验血液学杂志》2022年第5期1318-1323,共6页Journal of Experimental Hematology

基　　金：北京市科学技术委员会资助(Z171100001017103)。

摘　　要：目的:探讨lncRNA X染色体失活特异性转录因子(XIST)对急性髓系白血病细胞KG1a增殖和凋亡的影响及其相关作用机制。方法:收集北京航天中心医院2017年1月至2019年5月收治的41例急性髓系白血病患者,以及20例符合缺铁性贫血国际标准的患者作为对照组。将KG1a细胞分为pcDNA、pcDNA-XIST、pcDNA-XIST+miR-NC和pcDNA-XIST+miR-196b共4组。实时荧光定量PCR检测XIST和miR-196b表达水平,CCK-8检测细胞活性,流式细胞术检测细胞周期和细胞凋亡,Western blot法检测cleaved-caspase3、pro-caspase3、Bax、Bcl-2蛋白表达,双荧光素酶报告实验检测XIST和miR-196b的靶向关系。结果:急性髓系白血病组患者骨髓细胞中lncRNA XIST表达水平明显低于缺铁性贫血组(P<0.001)。KG1a细胞pcDNA-XIST组lncRNA XIST表达水平、G_(0)/G_(1)期细胞所占比例、细胞凋亡率及cleaved-caspase3和Bax的表达水平较pcDNA组均明显升高(均P<0.001),而miR-196b表达水平、细胞活性、S期细胞比例、pro-caspase3和Bcl-2表达水平均明显降低(均P<0.001)。pcDNA-XIST+miR-196b组细胞活性、S期细胞所占比例、pro-caspase3和Bcl-2表达水平较pcDNA-XIST组均明显升高(均P<0.001),而G_(0)/G_(1)期细胞所占比例、细胞凋亡率、cleaved-caspase3和Bax表达水平均明显降低(均P<0.001)。结论:过表达lncRNA XIST通过下调miR-196b表达抑制急性髓系白血病细胞KG1a增殖,促进细胞凋亡。Objective: To investigate the effect and molecular mechanism of lncRNA X-inactive specific transcript(XIST) on the proliferation and apoptosis of acute myeloid leukemia cells KG1a. Methods: Forty-one patients with acute myeloid leukemia from January 2017 to May 2019 treated in Beijing Aerospace Center Hospital were collected, as well as 20 patients who conformed to the international standard of iron deficiency anemia as control group. KG1a cells were divided into pcDNA group, pcDNA-XIST group, pcDNA-XIST+miR-NC group, and pcDNA-XIST+miR-196b group.Real-time fluorescence quantitative PCR was used to detect the expressions of XIST and miR-196b, CCK-8 was used to detect cell activity, flow cytometry was used to detect cell cycle and apoptosis, Western blot method was used to detect the protein expressions of cleaved-caspase3, pro-caspase3, Bax, and Bcl-2, and dual luciferase report experiment was used to detect the targeting relationship between XIST and miR-196b. Results: The expression level of lncRNA XIST in bone marrow cells in the AML group was significantly lower than that in the iron deficiency anemia group(P<0.001).Compared with pcDNA group, the expression level of lnc RNA XIST, proportion of cells in G_(0)/G_(1) phase, apoptosis rate, and the expression levels of cleaved-caspase3 and Bax in the pcDNA-XIST group of KG1a cells were significantly increased(all P<0.001), while the expression level of miR-196b, cell viability, the proportion of S-phase cells, and the expression levels of pro-caspase3 and Bcl-2 were significantly decreased(all P<0.001). Compared with pcDNA-XIST group, the cell activity, proportion of S-phase cells, and the expression levels of pro-caspase3 and Bcl-2 in the pcDNA-XIST+miR-196b group were significantly increased(all P<0.001), while the proportion of cells in the G_(0)/G_(1) phase, apoptosis rate, and the expression levels of cleaved-caspase3 and Bax decreased(all P<0.001). Conclusion: Overexpression of lncRNA XIST can inhibit the proliferation of acute myeloid leukemia cells KG1a a

关 键 词：lncRNA XIST miR-196b 急性髓系白血病 增殖 凋亡 

分 类 号：R733.7[医药卫生—肿瘤]