| 徐汉桥,方军,张梁.微 RNA-130a靶向人类 RUNT相关转录因子 3对肺癌 A549细胞增殖和转移的影响[J].安徽医药,2021,25(7):1396-1400. |
| 微 RNA-130a靶向人类 RUNT相关转录因子 3对肺癌 A549细胞增殖和转移的影响 |
| Effect of miR-130a targeting RUNX3 on proliferation and metastasis of lung cancer A549 cells |
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| DOI：10.3969/j.issn.1009-6469.2021.07.031 |
| 中文关键词: 癌，非小细胞肺 转录因子 3 微 RNA-130a 人类 RUNT相关转录因子 3 增殖 迁移 侵袭 上皮 -间质转化 通过， |
| 英文关键词: Carcinoma, non-small-cell lung Transcription factor 3 miR-130a RUNX3 Proliferation Migration Invasion Epithelial-mesenchymal transition |
| 基金项目: |
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| 摘要点击次数: 1996 |
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| 中文摘要: |
| 目的探讨微 RNA(miR)-130a与人类 RUNT相关转录因子 3(RUNX3)的靶向关系及其对非小细胞肺癌 A549细胞增殖、侵袭、迁移和上皮间质转化的影响。方法将体外培养的 A549细胞分为对照组(未处理)、anti-miR-NC组(转染 miR-130a inhibitor阴性对照)、 anti-miR-130a组(转染 miR-130a inhibitor)、 anti-miR-130a+siNC组(转染 miR-130a inhibitor和 NC-siRNA)和 antimiR-130a+siRUNX3组(转染 miR-130a inhibitor和 RUNX3-siRNA)，采用实时定量 PCR检测各组细胞中 miR-130a和 RUNX3mRNA的表达水平，蛋白质印迹法(Western Blot)检测各组细胞中 RUNX3蛋白和上皮型钙黏蛋白(E-cadherin)神经型钙黏蛋白(N-cadherin)和波形蛋白(Vimentin)的表达水平，四甲基偶氮唑盐微量酶反应比色法(MTT法)检测各组细胞增殖、能力，克隆形成实验检测各组细胞的克隆形成能力， Transwell小室实验检测各组细胞侵袭与迁移能力，荧光素酶实验检测 miR-130a和 RUNX3的靶向关系。结果荧光素酶实验证实， RUNX3是 miR-130a的潜在靶基因。与 anti-miR-NC组相比，下调 miR-130a可抑制 A549细胞增殖［(94.72±6.05)%比(51.28±3.24)%］、克隆［(36.15±2.06)%比( 15.46±1.25)%］侵袭［(90.25±5.48)个比( 42.18±3.26)个］、迁移［(118.55±9.86)个比(73.48±6.75)个］(P<0.05)，促进 E-cadherin蛋白表达［(0.26±03)比(0.48±0.04)］(P<0.05)而抑制 N-cadherin、Vimentin蛋白表达［(0.52±0.03)比( 0.25±0.02)；(0.68±0.05)比( 0.35±0.03)］(P<0.05)；与 anti-miR-130a+siNC相比，共转染 anti-miR-130a与 siRUNX3可成功增强 A549细胞增殖［(52.16±3.12)%比(72.23±6.13)%］、克隆［(16.08±1.05)%比.0、组，(22.12±1.34)%］、侵袭［(41.59±3.02)个比( 68.32±4.26)个］、迁移能力［(72.26±5.18)个比( 98.25±6.02)个］(P<0.05)抑制 E-cadherin蛋白表达［(0.52±0.04)比( 0.33±0.03)］(P<0.05)而促进 N-cadherin、Vimentin蛋白表达［(0.27±0.03)比( 0.46±0.03)；(0.33± .0.02)比(0.53±0.04)］(P<0.05)。结论 miR-130a可靶向 RUNX3抑制 A549细胞增殖、侵袭、迁移和上皮间质转移。 |
| 英文摘要: |
| Objective To investigate the targeting relationship between miR-130a and RUNX3 and its effects on proliferation, invasion, migration and epithelial-mesenchymal transition of lung cancer A549 cells.Methods A549 cells cultured in vitro were assigned into control group (untreated), anti-miR-NC group (transfected negative control of transfected miR-130a inhibitor), anti-miR-130a (transfected miR-130a inhibitor), anti-miR-130a+siNC (transfected miR-130a inhibitor and NC-siNC) and anti-miR-130a + siRUNX3 groups (transfected miR-130a inhibitor and RUNX3-siNC). The expression of miR-130a and RUNX3 mRNA in each group were detected by real-time quantitative PCR. Western Blot was used to detect the expression levels of RUNX3 protein and epithelial cadherin (Ecadherin), neural cadherin (N-cadherin) and vimentin (Vimentin) proteins in each group of cells. Cell proliferation abilities of eachgroup were checked by MTT method, the clone formation abilities of each group of cells were detected by clone formation, invasion andmigration abilities of each group were checked by Transwell lab assay. The targeting relationship between miR-130a and RUNX3 was detected by luciferase assay.Results Luciferase assay confirmed that RUNX3 was a potential target gene of miR-130a. Compared with the anti-miR-NC group, downregulation of miR-130a could inhibit the proliferation of A549 cells [(94.72±6.05)% vs. (51.28± 3.24)%], clone [(36.15±2.06)% vs. (15.46±1.25)% ], invasion [(90.25±5.48) vs. (42.18±3.26)], migration [(118.55±9.86) vs. (73.48± 6.75)] (P<0.05), promote the protein expression of E-cadherin [(0.26±0.03) vs. (0.48±0.04)] (P<0.05), while inhibit the expression of Ncadherin and Vimentin [(0.52±0.03) vs. (0.25±0.02); (0.68±0.05) vs. (0.35±0.03)] (P<0.05). Compared with the anti-miR-130a+siNC group, co-transfection of anti-miR-130a and siRUNX3 could successfully enhance the proliferation of A549 cells [(52.16±3.12)% vs. (72.23±6.13)%], clone [(16.08±1.05) )% vs. (22.12±1.34)%], invasion [(41.59±3.02) vs. (68.32±4.26)], migration ability [(72.26±5.18) vs. (98.25±6.02)] (P<0.05), inhibit the protein expression of E-cadherin [(0.52±0.04) vs. (0.33±0.03)] (P<0.05), and promote the protein expression of N-cadherin, Vimentin [(0.27±0.03) vs. (0.46±0.03); (0.33±0.02) vs. (0.53±0.04)] (P<0.05).Conclusion miR-130a can inhibit proliferation, invasion, migration and epithelial-mesenchymal metastasis of A549 cells by targeting RUNX3. |
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