| 鞠叶,秦世成,孙月,等.微 RNA-509-3p/干扰素刺激基因 15轴对喉癌细胞血管生成及血管内皮生长因子蛋白表达的影响[J].安徽医药,2024,28(8):1593-1598. |
| 微 RNA-509-3p/干扰素刺激基因 15轴对喉癌细胞血管生成及血管内皮生长因子蛋白表达的影响 |
| Effect of miR-509-3p/ISG15 axis on angiogenesis and VEGF protein expression in laryngeal cancer cells |
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| DOI：10.3969/j.issn.1009-6469.2024.08.023 |
| 中文关键词: 喉肿瘤 微 RNA-509-3p 干扰素刺激基因 15 血管生成 血管内皮生长因子 |
| 英文关键词: Laryngeal neoplasms MiR-509-3p ISG15 angiogenesis VEGF |
| 基金项目:山东省优秀中青年科学家科研奖励基金( BS2017SW13011) |
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| 中文摘要: |
| 目的探究微 RNA(miR)-509-3p/干扰素刺激基因 15(ISG15)轴对喉癌细胞血管生成及血管内皮生长因子(VEGF)蛋白表达的影响。方法 2022年 1―9月，将喉癌 M4E细胞分为 Con组、 miR-NC组、 miR-509-3p组、 pcDNA-ISG15组。逆转录聚合酶链式反应( RT-PCR)检测细胞中 miR-509-3p表达；细胞计数试剂盒( CCK8)检测细胞增殖能力； Transwell检测细胞侵袭能力；划痕实验检测细胞迁移能力；流式细胞仪检测细胞凋亡率； Matrigel体外成管实验检测血管生成能力；蛋白质印迹法检测细胞中 ISG15、VEGF蛋白表达；萤光素酶活性报告检测 miR-509-3p和 ISG15的靶向关系。结果与喉上皮细胞 NP69中 miR-509-3p表达(1.01±0.13)μg/L相比，喉癌细胞 SCC-40、SCC-2、M4E中 miR-509-3p表达［(0.61±0.08)(0.45±0.07)(0.18±0.07)μg/L］均降低，且 M4E细胞中 miR-509-3p表达低于 SCC-40、SCC-2细胞( P<0.05)。与 Con组相比， miR-50、9-3p组细胞中、miR-509-3p表达增加［( 1.17±0.05)μg/L比( 118±0.03)μg/L)］； Con组细胞中 miR-509-3p表达与 miR-NC组相比差异无统计学意义( P>0.05)。与 Con组相比， miR-509-3p组细胞增殖( 1.71±0.02比 3.09±0.07)、侵袭( 55.33±2.52比 150.67±2.52)、迁移( 30.58±2.08比 98.33±1.53)、形成小管数量( 61.28±2.15比 135.62±2.54)及细胞中 ISG15(0.24±0.03比 1.11±0.13)、 VEGF(0.35±0.02比 0.99±0.04)蛋白表达均降低，细胞凋亡率［( 18.76±0.18)%比( 5.61±0.08)%］增加( P<0.05)； Con组细胞增殖、侵袭、迁移、形成小管数量、凋亡率及细胞中 ISG15、VEGF蛋白表达和 miR-NC组相比差异无统计学意义( P>0.05)；与 miR-509-3p组相比， pcDNA-ISG15组细胞增殖( 3.25±0.06)、侵袭( 144.00±2.65)、迁移( 89.34±5.13)、形成小管数量( 131.56±2.08)及细胞中 ISG15(0.83±0.05)、 VEGF蛋白(0.89±0.03)表达增加，细胞凋亡率降低( 6.85±0.06)%(P<0.05)。结论过表达 miR-509-3p可抑制喉癌细胞血管生成及 VEGF蛋白表达。 |
| 英文摘要: |
| Objective To investigate the effect of miR-509-3p/ISG15 axis on angiogenesis and VEGF protein expression in laryngeal cancer cells.Methods The study was conducted between January 2022 and September 2022.The laryngeal carcinoma M4E cells weredivided into Con group, miR-NC group, miR-509-3p group, and pcDNA-ISG15 group. The rate of apoptosis; Matrigel in vitro tube for-mation assay to detect angiogenesis; protein immunoblotting to detect ISG15 and VEGF protein expression in cells; luciferase activityreporter to detect the targeting relationship between miR-509-3p and ISG15. Results Compared with the laryngeal epithelial cell NP69 (1.01±0.13)μg/L, the expression of miR-509-3p in laryngeal cancer cells SCC-40, SCC-2 and M4E was decreased (0.61±0.08, 0.45±0.07 and 0.18±0.07)μg/L, respectively. The expression of miR-509-3p in M4E cells was lower than that in SCC-40 and SCC-2 cells (P<0.05). Compared with the con group (118±0.03)μg/L, the expression of miR-509-3p in cells of the miR-509-3p group was in- creased (1.17±0.05)μg/L. The expression of miR-509-3p in Con group was not significantly different from that in miR-NC group (P> 0.05). Compared with con group [3.09±0.07, 150.67±2.52, 98.33±1.53, 135.62±2.54, (5.61±0.08)%], In miR-509-3p group, cell prolif-eration (1.71±0.02), invasion (55.33±2.52), migration (30.58±2.08), the number of tubules (1.28±2.15), ISG15 (0.24±0.03) and VEGF(0.35±0.02) protein expression in cells were decreased. The apoptosis rate (18.76±0.18)%,P<0.05) was increased.There was no signifi-cant difference in cell proliferation, invasion, migration, the number of tubules formed, apoptosis rate and the expression of ISG15 andVEGF protein in cells between the Con group and the miR-NC group (P>0.05). Compared with the miR-509-3p group,In the pcDNAISG15 group,the cell proliferation (3.25±0.06), invasion (144.00±2.65), migration (89.34±5.13), the number of tubules formed (131.56±2.08), and the expression of ISG15(0.83±0.05) and VEGF protein (0.89±0.03) in the cells increased the apoptosis rate was decreased(6.85±0.06)%(P<0.05).Conclusion Overexpression of miR-509-3p inhibits angiogenesis and VEGF protein expression in laryngeal cancer cells. |
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