目的探究miR-181a-5p对T淋巴细胞白血病(T lymphocytic leukemia, TLL)Jurkat细胞增殖及凋亡的影响。方法采用miR-181a-5p mimic、pc-B淋巴细胞瘤2(B-cell lymphoma 2, BCL-2)不同组合方式转染Jurkat细胞,并将细胞分为Jurkat组、mimic-scramble组、miR-181a mimic组、pc-BCL-2组和mimic+pc-BCL-2组。RT-PCR检测miR-181a-5p的表达及pc-BCL-2 mRNA水平,Western Blot检测BCL-2蛋白表达,CCK8和流式细胞学检测细胞增殖和凋亡情况。结果与mimic-scramble组比较,miR-181a mimic组miR-181a-5p表达水平显著升高,BCL-2 mRNA水平下降,差异有统计学意义( P <0.01)。与Jurkat组比较,miR-181a mimic组BCL-2蛋白表达量及细胞增殖倍数降低,细胞凋亡率明显升高,pc-BCL-2组BCL-2蛋白表达量及细胞增殖倍数升高,细胞凋亡率降低,差异有统计学意义( P <0.01);与miR-181a mimic组比较,mimic+pc-BCL-2组BCL-2蛋白表达量及细胞增殖倍数升高,细胞凋亡率显著下降,差异有统计学意义( P <0.01)。结论miR-181a-5p通过靶向抑制BCL-2表达以抑制TLL细胞系Jurkat细胞增殖、诱导细胞凋亡。
Objective To investigate the effects of miR-181a-5p on proliferation and apoptosis of T lymphocytic leukemia (TLL) cell line Jurkat. Methods Jurkat cells were transfected with different combinations of miR-181a 5p mimic and pc-B-cell lymphoma 2 (BCL-2) and divided into Jurkat group, mimic-scramble group, miR-182a mimic group, pc-BCL-2 group and mimic+pc-BCL-2 group. The expression of miR-181a-5p and mRNA levels of pc-BCL-2 were measured by RT-PCR, and protein level of BCL-2 was measured by Western Blot. In addition, CCK8 and flow cytometry were performed for cell proliferation and apoptosis. Results Compared with mimic-scramble group, the level of miR-181a-5p was up-regulated, while the level of BCL-2mRNA was down-regulated in miR-181a mimic group, and the difference is significant ( P <0.01). Compared with Jurkat group, the protein level of BCL-2 and proliferation folds were decreased markedly, while cell apoptosis rate was increased notably in miR-181a mimic group. In addition, the expression level of BCL-2 protein and proliferation folds were increased, whereas cell apoptosis rate was decreased significantly in pc-BCL-2 group, and there were significant differences ( P <0.01). Compared with miR-181a mimic group, the protein level of BCL-2 and proliferation folds were increased, while cell apoptosis was reduced in mimic+pc-BCL-2 group, suggesting significant differences ( P <0.01). Conclusion miR-181a-5p inhibits cell proliferation and induces apoptosis of TLL cell line Jurkat by targeting inhibition of BCL-2 expression.
Clinical Misdiagnosis & Mistherapy