| 张传政,常欣辛,张晓东,等.微小 RNA-646对胶质母细胞瘤细胞系 U251增殖和转移的影响[J].安徽医药,2023,27(1):148-153. |
| 微小 RNA-646对胶质母细胞瘤细胞系 U251增殖和转移的影响 |
| Effect of miR-646 on proliferation and metastasis of glioblastoma cell line U251 |
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| DOI：10.3969/j.issn.1009-6469.2023.01.033 |
| 中文关键词: 胶质母细胞瘤 微小 RNA-646 细胞增殖 转移 叉头框蛋白 K1 |
| 英文关键词: Glioblastoma miR-646 Cell proliferation Metastasis FOXK1 |
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| 中文摘要: |
| 目的探讨微小 RNA-646(miR-646)对胶质母细胞瘤细胞系 U251增殖和转移的影响及机制。方法 2014年 1月至 2016年 1月，从成都市第六人民医院采集 14对神经胶质瘤病人肿瘤组织样本和相邻的非肿瘤样本，并于 2018年 5月至 2019年 5月，体外培养正常星形胶质细胞 HNAs和人胶质母细胞瘤细胞系 SHG44、A172和 U251，实时荧光定量 PCR检测 miR-646在胶质瘤组织、胶质瘤细胞 SHG44、A172和 U251中的表达。将体外培养的 U251细胞分为模拟物对照组、 miR-646模拟物组、抑制剂对照组、 miR-646抑制剂组、 miR-646模拟物 +叉头框 K1过表达质粒(pcDNA-FOXK1)组和 miR-646模拟物 +pcDNA组，采用实时荧光定量 PCR检测细胞中 miR-646表达，检测细胞增殖、侵袭和迁移能力，蛋白质印迹法( Western blotting)检测细胞中叉头框 K1(FOXK1)和上皮间质转化相关蛋白表达。双萤光素酶报告基因实验检测 miR-646与 FOXK1的靶向关系。结果与正常脑组织相比，胶质瘤组织中 miR-646表达水平( 0.36±0.03比 1.00±0.06)显著降低( P<0.05)；与正常星形胶质细胞相比，胶质瘤细胞 SHG44、A172和 U251中 miR-646表达水平( 0.54±0.04、0.63±0.06、0.38±0.04比 1.00±0.08)显著降低( P<0.05)。与模拟物对照组相比，转染 miR-646模拟物可明显上调 U251细胞中 miR-646表达( 5.28±0.53比 1.00±0.11)和上皮钙黏素( E-cadherin)蛋白水平，减弱细胞增殖、克隆形成、侵袭和迁移能力，抑制 FOXK1(0.28±0.03比 0.51±0.04)、神经钙黏素( N-cadherin)和波形蛋白( Vimentin)蛋白表达( P<0.05)；与抑制剂对照组相比，转染 miR-646抑制剂可得到与 miR-646模拟物处理相反的结果( P< |
| 英文摘要: |
| Objective To explore the effect and mechanism of miR-646 on proliferation and metastasis of glioblastoma cell line U251.Method Fourteen pairs of tumor tissue samples and adjacent non-tumor samples from patients with glioma in Chengdu SixthPeople′s Hospital from January 2014 to January 2016 were collected. Normal astrocytes HNAs and human glioblastoma cell linesSHG44, A172 and U251 were cultured in vitro from May 2018 to May 2019. Real-time fluorescent quantitative PCR was used to detect the expression of miR-646 in glioma tissue, glioma cells SHG44, A172, and U251. U251 cells cultured in vitro were assigned into mock control group, miR-646 mimic group, inhibitor control group, miR-646 inhibitor group, miR-646 mimic + forkhead box K1 overexpression plasmid (pcDNA-FOXK1) group and miR-646 mimic + pcDNA group. The expression of miR-646 was detected by real-time fluorescent quantitative PCR, and the ability of cell proliferation, invasion and migration were detected. The expression of forkhead box K1(FOXK1) and epithelial mesenchymal transformation related proteins in cells was detected by western blotting. The dual luciferase reporter gene assay was used to detect the targeting relationship between miR-646 and FOXK1.Result Compared with normal brain tissue, the expression level of miR-646 in glioma tissue (0.36±0.03 vs. 1.00±0.06) was significantly decreased (P<0.05). Compared with normal astrocytes, the expression of miR-646 in glioma cells SHG44, A172 and U251 (0.54±0.04, 0.63±0.06, 0.38±0.04 vs. 1.00±0.08) was significantly reduced (P<0.05). Compared with the mock control group, transfection of miR-646 mimic significantly increased the expression of miR-646 (5.28±0.53 vs. 1.00±0.11) and E-cadherin protein in U251 cells, attenuated cell proliferation, cell clone formation, invasion and migration, and inhibited the expression of FOXK1 (0.28±0.03 vs. 0.51±0.04), N-cadherin and Vimentin (P<0.05); the transfection of miR-646 inhibitor resulted in the opposite result of miR-646 mimic treatment compared with the inhibitor control group (P<0.05). Compared with miR-646 mimic + pcDNA group, miR-646 mimic + pcDNA-FOXK1 group significantly improved the proliferation, clonal formation, invasion and migration ability of U251 cells (P<0.05).Conclusion miR-646 can inhibit the proliferation andmetastasis of U251 cells, and its mechanism may be related to the targeted regulation of FOXK1 expression. |
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