| 刘莹,连海伟,易伟,等.环状 RNA同源性蛋白激酶 3靶向微 RNA-338促进胶质瘤细胞侵袭、迁移的实验研究[J].安徽医药,2024,28(1):138-142. |
| 环状 RNA同源性蛋白激酶 3靶向微 RNA-338促进胶质瘤细胞侵袭、迁移的实验研究 |
| Experimental study on CircHIPK3 targeting mir-338 promoting invasion and migration of glioma cells |
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| DOI：10.3969/j.issn.1009-6469.2024.01.029 |
| 中文关键词: 神经胶质瘤 微 RNA-338 人血清环状 RNA同源性蛋白激酶 3 迁移 侵袭 U251细胞 |
| 英文关键词: Glioma MicroRNA-338 Human serum cyclic RNA homologous protein kinase 3 Migration Invasion U251cells |
| 基金项目:湖北省自然科学基金( 2020CFB256) |
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| 中文摘要: |
| 目的探讨人血清环状 RNA同源性蛋白激酶 3(CircHIPK3)靶向微 RNA-338(miR-338)对胶质瘤细胞 U251细胞侵袭、迁移的影响。方法 2021年 2—12月，在武汉大学人民医院科研中心将 U251细胞分为空白( NG)组、 CircHIPK3阴性对照( shcontrol)组、 HIPK3敲减( sh-CircHIPK3)组，实时荧光定量 PCR(qRT-PCR)检测 U251细胞中 CircHIPK3、miR-338表达水平； Tran- swell检测细胞迁移与侵袭；划痕法检测细胞迁移；流式细胞术检测细胞周期；通过 Circular RNA Interactome、RegRNA2.0、Circ- Bank Database网站预测 CircHIPK3(ID:hsa_circ_0000284)的靶向 miRNA并用双萤光素酶实验验证，蛋白质印迹法检测基质金属蛋白酶( MMP)-2、MMP-9蛋白表达。结果与 NG组、 sh-control组比较， sh-CircHIPK3组中 CircHIPK3(1.00±0.00、1.06±0.26比 0.56±0.06)表达水平显著降低( P<0.05)miR-338(1.00±0.00、1.12±0.19比 1.89±0.28)表达、 G1期细胞比例［( 58.72±0.36)%、(58.45±0.27)%比( 64.72±0.47)%］升高( P<005)U251细胞侵袭数目［( 164.89±12.55)个、(165.77±12.16)个比( 80.13±11.37)个］、划痕愈合率［( 25.66±2.37)%、(26.38±2.53(10.36±1.53)%］、迁移细胞数目［( 196.72±18.75)个、(194.65±17.86)个比)%比，(95.58±8.66)个］、 S期细胞比例［( 26.45±0.39)%、(26.57±0.41)%比( 20.72±0.18)%］明显降低( P<0.05)； miR-338是 CircHIPK3的靶基因。与 NG组、 sh-control组比较， sh-CircHIPK3组 MMP-2(1.31±0.23、1.33±0.20比 0.61±0.05)、 MMP-9(1.16±0.22、1.15±0.21比 0.85±0.19)蛋白表达水平均显著降低( P<0.05)。结论沉默 CircHIPK3通过靶向上调 miR-338表达能抑制胶质瘤细胞 U251细胞迁移和侵袭。 |
| 英文摘要: |
| Objective To investigate the effect of human serum cyclic RNA homologous protein kinase 3 (CircHIPK3) targeting mi- croRNA-338 (miR-338) on the migration and invasion of glioma cells U251.Methods Glioma cells U251 were assigned into blank (NG) group, CircHIPK3 negative control (sh-control) group and HIPK3 knockdown (sh-CircHIPK3) group at the Scientific ResearchCenter of the People's Hospital of Wuhan University from February to December 2021. The expression levels of CircHIPK3 and mir338 in U251 cells were detected by real-time PCR (qRT-PCR); the migration and invasion ability of cells were detected by Transwell;the migration ability of cells was detected by scratch method; the cell cycle was detected by flow cytometry; CircHIPK3 was predictedby circular RNA interactome, RegRNA2.0 and CircBank Database website (ID: hsa_circ_0000284) and verified by double luciferase experiment. The expression of matrix metalloproteinase (MMP)-2 and MMP-9 protein were detected by Western blotting.Results Com- pared with NG group and sh-control group, the expression level of CircHIPK3 (1.00±0.00, 1.06±0.26 vs. 0.56±0.06) in sh-CircHIPK3 group was decreased significantly (P< 0.05); the expression of mir-338 (1.00±0.00, 1.12±0.19 vs. 1.89±0.28) and the proportion of cells in G1 phase [(58.72±0.36)% , (58.45±0.27)% vs. (64.72±0.47)% ] increased significantly (P<0.05), the number of invasion (164.89± 12.55, 165.77±12.16 vs. 80.13±11.37) and scratch healing rate [(25.66±2.37)%, (26.38±2.53)% vs. (10.36±1.53)%], the number of mi- grating cells (196.72±18.75, 194.65±17.86 vs. 95.58±8.66), the proportion of cells in S phase [(26.45±0.39)% , (26.57±0.41)% vs. (20.72±0.18)%] of U251 cells were significantly decreased (P<0.05); mir-338 was target gene of CircHIPK3. Compared with NG group and sh-control group, the expression levels of MMP-2 (1.31±0.23, 1.33±0.20 vs. 0.61±0.05) and MMP-9 (1.16±0.22, 1.15±0.21 vs. 0.85±0.19) in the sh-CircHIPK3 group were significantly decreased (P<0.05).Conclusion Silencing CircHIPK3 inhibits glioma cell U251 cell migration and invasion by targeting the up-regulation of miR-338 expression. |
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