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| circUBE2D2 靶向 miR-376a-3p 调控结直肠癌 SW620 细胞的增殖、迁移及侵袭 | |
| 引用本文： | 李明辉,王裕,白晓刚. circUBE2D2 靶向 miR-376a-3p 调控结直肠癌 SW620 细胞的增殖、迁移及侵袭[J]. 医学分子生物学杂志, 2022, 19(4): 326-331. DOI: 10.3870/j.issn.1672-8009.2022.04.010 |
| 作者姓名： | 李明辉 王裕 白晓刚 |
| 作者单位： | 大连大学附属新华医院肛肠外科 辽宁省大连市, 116011 |
| 摘 要： | 目的 探讨 circUBE2D2 对结直肠癌 SW620 细胞增殖、 迁移及侵袭的影响及其可能作用机制。 方法 收集 2020 年 1 月至 2020 年 5 月大连大学附属新华医院肛肠外科收治的 45 例结直肠癌患者的癌组织及其相应癌旁组织标本, 采用 qRT-PCR 法检测 circUBE2D2、 miR-376a-3p 的表达量; 以人结直肠癌细胞SW620 为研究对象, 随机分为 si-NC 组、 si-circUBE2D2 组、 miR-NC 组、 miR-376a-3p 组、 si-circUBE2D2 +anti-miR-NC 组和 si-circUBE2D2 + anti-miR-376a-3p 组; CCK-8 法、 平板克隆形成实验、 划痕实验与 Transwell 实验分别检测细胞增殖、 克隆形成、 迁移及侵袭; 双荧光素酶报告实验检测 miR-376a-3p 过表达对野生型载体 WT-circUBE2D2 的荧光素酶活性; Western 印迹法检测上皮型钙黏蛋白 (E-cadherin)、 神经型钙黏蛋白 (N-cadherin) 蛋白表达量。 结果 与癌旁组织比较, 结直肠癌组织中 circUBE2D2 的表达量升高(P< 0. 05), miR-376a-3p 的表达量降低 (P< 0. 05); 与 si-NC 组比较, si-circUBE2D2 组细胞活力、 划痕愈合率和 N-cadherin 蛋白水平降低 (P< 0. 05), 细胞克隆形成数和侵袭细胞数减少 (P< 0. 05), E-cadherin 蛋白水平升高 (P< 0. 05); 与 miR-NC 组比较, miR-376a-3p 组细胞活力、 划痕愈合率和 N-cadherin 蛋白水平降低 (P< 0. 05), 细胞克隆形成数和侵袭细胞数减少 (P< 0. 05), E-cadherin 蛋白水平升高 (P< 0. 05);miR-376a-3p 过表达可抑制野生型载体 WT-circUBE2D2 的荧光素酶活性 (P< 0. 05); 与 si-circUBE2D2 + anti-miR-NC 组比较, si-circUBE2D2 + anti-miR-376a-3p 组细胞活力、 划痕愈合率和 N-cadherin 蛋白水平升高(P< 0. 05), 细胞克隆形成数和侵袭细胞数增多 (P< 0. 05), E-cadherin 蛋白水平降低 (P< 0. 05)。 结论 干扰 circUBE2D2 表达可通过靶向调控 miR-376a-3p 表达而降低结直肠癌细胞增殖、 克隆形成、 迁移及侵袭能力。 |
| 关 键 词： | 结直肠癌 circUBE2D2 miR-376a-3p 细胞增殖 迁移 侵袭 |
circUBE2D2 Regulates Proliferation,Migration and Invasion ofSW620 Colorectal Cancer Cells via miR-376a-3p |
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| LI Minghui,WANG Yu,BAI Xiaogang. circUBE2D2 Regulates Proliferation,Migration and Invasion ofSW620 Colorectal Cancer Cells via miR-376a-3p[J]. Journal of Medical Molecular Biology, 2022, 19(4): 326-331. DOI: 10.3870/j.issn.1672-8009.2022.04.010 | |
| Authors: | LI Minghui WANG Yu BAI Xiaogang |
| Affiliation: | Department of Anorectal Surgery, Xinhua Hospital Affiliated to Dalian University, Dalian, Liaoning, 116011, China |
| Abstract: | Objective To explore the effect of circUBE2D2 on the proliferation, migration andinvasion of SW620 colorectal cancer cells and its underlying mechanism. Methods The cancer tissues and the corresponding para-cancerous tissues from 45 patients with colorectal cancer admitted toour hospital from January 2020 to May 2020 were collected. The expression levels of circUBE2D2and miR-376a-3p were detected by qRT-PCR. Human colorectal cancer cell line SW620 was used inthis research and were divided into groups as follows: si-NC group, si-circUBE2D2 group, miR-NC group, miR-376a-3p group, si-circUBE2D2 + anti-miR-NC group, and si-circUBE2D2 + anti-miR-376a-3p group. The CCK-8 and plate colony formation assay were used to measure the proliferation and colony formation of cells. The migration and invasion of cells were determined by the woundhealing assay and the transwell assay. The dual luciferase reporter experiment was used to assay theeffect of miR-376a-3p overexpression on the luciferase activity of the wild-type vector WT-circUBE2D2. The expression levels of E-cadherin and N-cadherin proteins were detected by Westernblotting. Results The expression level of circUBE2D2 was increased in colorectal cancer tissues,while that of miR-376a-3p was decreased when compared with adjacent tissues (P< 0. 05). The cellviability and scratch healing rate were reduced, the number of colonies formed, the number of invasive cells and the expression level of N-cadherin were decreased in the si-circUBE2D2 group and themiR-376a-3p group when compared with the si-NC group and the miR-NC group, while the expression level of E-cadherin was increased respectively ( P < 0. 05 ). Overexpression of miR-376a-3pcould inhibit the luciferase activity of wild-type vector WT-circUBE2D2 (P< 0. 05). The cell viability, the scratch healing rate, the number of cell colony formed, the number of invasive cells, andthe expression level of N-cadherin were increased in the si-circUBE2D2 + anti-miR-376a-3p group,while the expression level of E-cadherin was decreased when compared with the si-circUBE2D2 +anti-miR-NC group (P< 0. 05). Conclusion Interference of circUBE2D2 expression could reducethe proliferation, colony formation, migration and invasion of colorectal cancer cells by targetingmiR-376a-3p. |
| Keywords: | colorectal cancer circUBE2D2 miR-376a-3p cell proliferation migration invasion |
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