| lncRNA MAGI2-AS3下调miR-31-5p抑制肺腺癌细胞的增殖、迁移、侵袭并促进凋亡 |
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| 引用本文： | 龚年金,李光才,张明华,刘培俊,向悦华. lncRNA MAGI2-AS3下调miR-31-5p抑制肺腺癌细胞的增殖、迁移、侵袭并促进凋亡[J]. 中华细胞与干细胞杂志(电子版), 2020, 10(4): 204-212. DOI: 10.3877/cma.j.issn.2095-1221.2020.04.002 |
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| 作者姓名： | 龚年金 李光才 张明华 刘培俊 向悦华 |
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| 作者单位： | 1. 445000 恩施，湖北省恩施土家族苗族自治州中心医院呼吸内科 |
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| 摘 要： | 目的研究lncRNA MAGI2-AS3对肺癌A549细胞增殖、迁移、侵袭和凋亡的影响和潜在的分子机制。方法根据转染载体不同将A549细胞分为pcDNA3.1组(转染pcDNA3.1)、pcDNA3.1-MAGI2-AS3组(转染pcDNA3.1-MAGI2-AS3)、anti-miR-NC组(转染anti-miR-NC)、anti-miR-31-5p组(转染anti-miR-31-5p)、pcDNA3.1-MAGI2-AS3+miR-NC组(共转染pcDNA3.1-MAGI2-AS3和miR-NC)、pcDNA3.1-MAGI2-AS3+miR-31-5p组(共转染pcDNA3.1-MAGI2-AS3和miR-31-5p mimics)。实时荧光定量PCR(qRT-PCR)检测miR-31-5p和MAGI2-AS3 RNA的表达,四氮唑蓝(MTT)法测定A549细胞增殖活性,Transwell实验检测细胞迁移和侵袭能力,双荧光素酶报告系统验证MAGI2-AS3与miR-31-5p的调控关系,流式细胞术检测细胞凋亡与周期。两组间比较采用独立样本t检验进行分析;多组间比较采用单因素方差分析,组内多重比较采用SNK-q检验。结果与人正常肺细胞HBE相比,肺癌细胞A549中的MAGI2-AS3表达量(0.48±0.03比1.29±0.06)降低,miR-31-5p表达量(1.01±0.05比0.25±0.02)升高;与pcDNA3.1组比较,pcDNA3.1-MAGI2-AS3组A549细胞活力(0.48±0.04比0.77±0.06)、迁移[(81.33±2.87)个比(124.33±3.09)个]和侵袭[(32.00±2.83)个比(53.00±3.27)个]细胞数、S期细胞所占比例(23.01﹪±1.00﹪比32.95﹪±1.06﹪)均降低,凋亡率(19.95﹪±1.25﹪比7.23﹪±0.51﹪)、G0-G1期细胞所占比例(43.58﹪±2.15﹪比33.56﹪±1.23﹪)均升高;与anti-miR-NC组比较,anti-miR-31-5p组A549细胞活力(0.53±0.04比0.78±0.06)、迁移[(76.00±3.74)个比(108.33±2.87)个]和侵袭[(30.00±1.63)个比(42.33±2.05)个]细胞数、S期细胞所占比例(24.43﹪±1.13﹪比32.91﹪±1.08﹪)降低,凋亡率(18.21﹪±1.24﹪比7.29﹪±0.51﹪)、G0-G1期细胞所占比例(41.56﹪±2.19﹪比33.53﹪±1.27﹪)升高,差异有统计学意义(P均<0.05);双荧光素酶报告系统结果显示,MAGI2-AS3靶向负调控miR-31-5p的表达。与pcDNA3.1-MAGI2-AS3+miR-NC组比较,pcDNA3.1-MAGI2-AS3+miR-31-5p组A549细胞活力(0.68±0.06比0.50±0.04)、迁移[(91.00±1.63)个比(52.67±2.62)个]和侵袭[(62.67±2.49)个比(31.67±4.03个)]细胞数升高,凋亡率(10.59﹪±1.0﹪比21.11﹪±1.14﹪)降低,差异有统计学意义(P均<0.05)。结论lncRNA MAGI2-AS3通过靶向miR-31-5p抑制A549细胞的增殖、迁移和侵袭,促进细胞凋亡。lncRNA MAGI2-AS3是肺癌潜在分子治疗靶点。
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| 关 键 词： | 肺癌 A549细胞 MAGI2-AS3 MiR-31-5p 增殖 迁移 侵袭 凋亡 |
| 收稿时间： | 2019-10-17 |
Effects of lncRNA MAGI2-AS3 targeting miR-31-5p on the inhibition of proliferation,migration,invasion and promotion of apoptosis of lung adenocarcinoma cancer cells |
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| Nianjin Gong,Guangcai Li,Minghua Zhang,Peijun Liu,Yuehua Xiang. Effects of lncRNA MAGI2-AS3 targeting miR-31-5p on the inhibition of proliferation,migration,invasion and promotion of apoptosis of lung adenocarcinoma cancer cells[J]. , 2020, 10(4): 204-212. DOI: 10.3877/cma.j.issn.2095-1221.2020.04.002 |
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| Authors: | Nianjin Gong Guangcai Li Minghua Zhang Peijun Liu Yuehua Xiang |
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| Affiliation: | 1. Department of Respiratory Medicine, Enshi Tujia and Miao Autonomous Prefecture Central Hospital, Enshi 445000, China |
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| Abstract: | ObjectiveTo investigate the effects and mechanisms of lncRNA MAGI2-AS3 on the proliferation, migration, invasion and apoptosis of lung cancer A549 cells. MethodsA549 cells were divided into pcDNA3.1 group (transfected with pcDNA3.1), pcDNA3.1-MAGI2-AS3 group (transfected with pcDNA3.1-MAGI2-AS3), anti-miR-NC group (transfected with anti-miR-NC), anti-miR-31-5p group (transfected with anti-miR-31-5p), pcDNA3.1-MAGI2-AS3 + miR-NC group (co-transfected with pcDNA3.1-MAGI2-AS3 and miR-NC), pcDNA3.1-MAGI2-AS3 + miR-31-5p group (co-transfection with pcDNA3.1-MAGI2-AS3 and miR-31-5p mimics). The RNA expression levels of miR-31-5p and MAGI2-AS3 were detected by qRT-PCR. The proliferation rate of A549 cells was measured by MTT assay. Migration and invasion abilities of A549 cells were determined by Transwell assay. The relationship between lncRNA MAGI2-AS3 and miR-31-5p was verified by dual-luciferase reporter assay system. Apoptosis and cell cycle of A549 cells were detected by flow cytometry. The differences between groups were compared by t-test, and the differences among groups were compared by ANOVA and SNK-q test. ResultsCompared with normal lung cell HBE, the expression levels of MAGI2-AS3 in lung cancer cell A549 was significantly decreased (0.48±0.03 vs 1.29±0.06), and the expression levels of miR-31-5p was remarkably increased (1.01±0.05 vs 0.25±0.02). Compared with pcDNA3.1 group, the viability (0.48±0.04 vs 0.77±0.06), the cell numbers of migration (81.33±2.87 vs 124.33±3.09) and invasion (32.00±2.83 vs 53.00±3.27), proportion of S-phase cells (23.01﹪± 1.00﹪ vs 32.95﹪± 1.06﹪) in A549 cells of pcDNA3.1-MAGI2-AS3 group were significantly reduced, while the apoptosis rate (19.95﹪± 1.25﹪ vs 7.23﹪± 0.51﹪) and proportion of cells in G0-G1 phase (43.58﹪± 2.15﹪ vs 33.56﹪± 1.23﹪) were significantly increased. Compared with anti-miR-NC group, the viability (0.53±0.04 vs 0.78±0.06), the cell numbers of migration (76.00±3.74 vs 108.33±2.87) and invasion (62.67± 2.49 vs 31.67±4.03), proportion of cells in S phase (24.43﹪±1.13﹪ vs 32.91﹪±1.08﹪) in A549 cells of anti-miR-31-5p group were significantly reduced, while the apoptosis rate (18.21﹪ ± 1.24﹪ vs 7.29﹪±0.51﹪) and proportion of cells in G0-G1 phase (41.56﹪± 2.19﹪vs 33.53﹪ ± 1.27﹪) were significantly increased, and the difference is statistically significant (all P < 0.05). The results of dual-luciferase reporter assay system showed that MAGI2-AS3 targeted and negatively regulated the expression of miR-31-5p. Compared with pcDNA3.1-MAGI2-AS3+miR-NC group, the viability (0.68±0.06 vs 0.50±0.04), the cell numbers of migration (91.00±1.63 vs 52.67 ± 2.62) and invasion (62.67 ± 2.49 vs 31.67 ± 4.03) in A549 cells of pcDNA3.1-MAGI2-AS3+miR-31-5p group were significantly increased, and the apoptosis rate (10.59﹪±1.01﹪vs 21.11﹪±1.14﹪) was decreased significantly, and the difference is statistically significant (all P < 0.05) . ConclusionsLncRNA MAGI2-AS3 could inhibite the proliferation, migration and invasion of A549 cells and promoted cell apoptosis by targeting miR-31-5p. LncRNA MAGI2-AS3 is a potential molecular therapeutic target for lung cancer. |
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| Keywords: | Lung cancer A549 cell MAGI2-AS3 miR-31-5p Proliferation Migration Invasion Apoptosis |
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