| 杨莉,刘炜,李彦格,等.微小RNA-150 靶向c-Myb 表达对T 淋巴细胞白血病Jurkat细胞增殖的影响[J].安徽医药,2022,26(8):1615-1618. |
| 微小RNA-150 靶向c-Myb 表达对T 淋巴细胞白血病Jurkat细胞增殖的影响 |
| MiR-150 targeting c-Myb expression inhibits the proliferation of Jurkat cells |
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| DOI：10.3969/j.issn.1009-6469.2022.08.030 |
| 中文关键词: 白血病，T细胞 微小RNA-150 c-Myb Jurkat细胞 细胞增殖 |
| 英文关键词: Leukemia,T-Cell microRNA-150 c-Myb Jurkat cell Cell proliferation |
| 基金项目:2018年度河南省医学科技攻关计划(2018020679) |
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| 中文摘要: |
| 目的探讨微小RNA-150(miR-150)靶向c-Myb表达对人T淋巴细胞白血病(T-LL)Jurkat细胞增殖的影响。方法体外培养人Jurkat细胞，分为空白对照组(不转染)、阴性对照组(转染negative control-miR-150)和miR-150过表达组(转染hsamiR-150 mimic)。实时荧光定量PCR(qRT-PCR)法检测Jurkat细胞中miR-150 及c-Myb 表达；人胆囊收缩素/缩胆囊素八肽(CCK-8)法检测Jurkat细胞增殖；流式细胞仪检测各组Jurkat细胞凋亡情况；双荧光素酶报告基因实验检测miR-150和c-Myb的靶向关系；蛋白质印迹法(Western blotting)检测各组Jurkat细胞中c-Myb、增殖细胞核抗原(PCNA)、Bcl-2相关X蛋白(Bax)蛋白表达情况。结果空白对照组与阴性对照组Jurkat细胞中miR-150表达水平、c-Myb mRNA及蛋白表达水平、OD值、细胞凋亡率、及PCNA、Bax蛋白水平差异无统计学意义(P>0.05)；与空白对照组相比，miR-150过表达组Jurkat细胞中miR-150表达水平［(1.42±0.14)比(1.03±0.09)］、凋亡率［(20.79±1.15)%比(8.76±0.74)%］、Bax蛋白表达水平［(0.65±0.18)比(0.42±0.13)］显著升高(P<0.05)，c-Myb mRNA［(0.66±0.14)比(0.98±0.09)］及蛋白表达水平［(0.37±0.06)比(0.64±0.12)］、OD值［(0.25±0.06)比(0.46±0.09)］、PCNA蛋白表达水平［(0.33±0.07)比(0.56±0.11)］显著降低(P<0.05)。双荧光素酶报告基因实验显示，c-Myb是miR-150的靶基因。结论miR-150靶向c-Myb表达抑制Jurkat细胞增殖并诱导其凋亡。 |
| 英文摘要: |
| Objective To investigate the effect of microRNA-150 (miR-150) targeting c-Myb expression on the proliferation of Jur?kat cells.Methods Human Jurkat cells were cultured in vitro and assigned into blank control group (no transfection), negative control group (transfected with negative control-miR-150) and miR-150 overexpression group (transfected with hsa-miR-150 mimic). The ex?pressions of miR-150 and c-Myb in Jurkat cells were detected by real-time fluorescent quantitative PCR (qRT-PCR); the proliferation of Jurkat cells was detected by cholecystokinin octapeptide (CCK-8); the apoptosis of Jurkat cells was detected by flow cytometry; the tar?geting relationship between miR-150 and c-Myb was detected by double luciferase reporter gene assay; and Western blotting (WB) was used to detect the protein expressions of c-Myb, proliferating cell nuclear antigen (PCNA) and Bcl-2-associated X-protein Bax in Jurkat cells.Results There were no significant differences in the expression levels of miR-150, c-Myb mRNA and protein, OD value, apopto?sis rate, protein expression levels of PCNA and Bax in Jurkat cells between blank control group and negative control group (P>0.05).Compared with blank control group, the expression level of miR-150 [(1.42±0.14) vs. (1.03±0.09)], apoptosis rate [(20.79±1.15) % vs.(8.76±0.74) %] and Bax protein [(0.65±0.18) vs. (0.42±0.13)] in Jurkat cells of miR-150 overexpression group were significantly higher(P<0.05), while the expression levels of c-Myb mRNA [(0.66±0.14) vs. (0.98±0.09)] and protein [(0.37±0.06) vs. (0.64±0.12)], OD value [(0.25±0.06) vs. (0.46±0.09)] and protein expression level of PCNA [(0.33±0.07) vs. (0.56±0.11)] were significantly lower (P<0.05). Dou?ble luciferase reporter gene experiment results showed that c-Myb was the target gene of miR-150.Conclusion The miR-150 targeting c-Myb expression inhibits the proliferation and induces the apoptosis of Jurkat cells. |
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