| 鲁艳妮,焦琳,周清文,等.微小 RNA-4478下调鼠双微体 -2对结肠癌细胞增殖、迁移、侵袭的影响[J].安徽医药,2021,25(5):938-942. |
| 微小 RNA-4478下调鼠双微体 -2对结肠癌细胞增殖、迁移、侵袭的影响 |
| Effect of miR-4478 down-regulating MDM2 on proliferation, migration and invasion of colon cancer cells and its molecular mechanism |
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| DOI：10.3969/j.issn.1009-6469.2021.05.023 |
| 中文关键词: 结肠肿瘤 微小 RNA-4478 鼠双微体 -2 增殖 迁移 侵袭 |
| 英文关键词: Colonic neoplasms miR-4478 MDM2 Proliferation Migration Invasion |
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| 中文摘要: |
| 目的探讨微小 RNA-4478(miR-4478)对结肠癌细胞增殖、迁移、侵袭的作用并阐明相关机制。方法实时定量聚合酶链式反应( PCR)检测 miR-4478在结肠癌组织中的表达。结肠癌 SW1116细胞转染 miR-4478 mimics及阴性对照后，分别采用 3(-4，5-二甲基噻唑 -2)-2，5-二苯基四氮唑溴盐( MTT)和小室( Transwell)法检测细胞的增殖、迁移及侵袭。预测 miR-4478的靶基因，双荧光素酶报告基因实验验证靶基因。结肠癌 SW1116细胞转染 miR-4478 mimics或抑制物后，蛋白质免疫印迹(Western blot)法检测细胞靶基因表达。靶基因过表达验证细胞增殖、迁移及侵袭。 Western blot法检测细胞周期蛋白 D1(Cyclin D1)、基质金属蛋白酶 2(MMP-2)、 P21、钙粘附蛋白 E(E-cadherin)蛋白表达。结果相对于癌旁组织， miR-4478在结肠癌组织中的表达显著降低［(0.84±0.07)比( 0.26±0.02)t=48.461，P<0.05］；转染 miR-4478 mimics可显著抑制细胞增殖、迁移及侵袭(P<0.001)。 miR-4478可负向调控 MDM2表达( P<0.，001)。 miR-4478过表达可抑制由 MDM2过表达导致的结肠癌细胞增殖、迁移及侵袭(P<0.001)。结论 miR-4478通过抑制 MDM2表达而抑制结肠癌细胞增殖、迁移及侵袭。 |
| 英文摘要: |
| Objective To investigate the effects of microRNA-4478 (miR-4478) on proliferation, migration and invasion of colon cancer cells and to elucidate the relevant mechanisms.Methods Real-time quantitative PCR was used to detect the expression of miR4478 in colon cancer tissues. Colon cancer SW1116 cells were transfected with miR-4478 mimics and negative controls, and cell proliferation, migration and invasion were detected by MTT and Transwell methods, respectively. The target gene of miR-4478 was predicted, and the dual luciferase reporter gene experiment was used to verify the target gene. After colon cancer SW1116 cells were transfected with miR-4478 mimics or inhibitors, the expression of target genes was detected by Western blot. Target gene overexpression confirmed cell proliferation, migration and invasion. The expressions of Cyclin D1, MMP-2, P21 and E-cadherin were detected by Western blot.Results The expression of miR-4478 was significantly reduced in colon cancer tissues[ (0.84±0.07) vs. (0.26±0.02),t=48.461,P< 0.05]. Transfection of miR-4478 mimics significantly inhibited cell proliferation, migration and invasion(P<0.001). miR-4478 could negatively regulate MDM2 expression (P<0.001). Overexpression of miR-4478 inhibited colon cancer cell proliferation, migration and invasion caused by overexpression of MDM2(P<0.001).Conclusion miR-4478 inhibits colon cancer cell proliferation, migration and invasion by inhibiting MDM2 expression. |
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