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本研究初步探讨过表达miRNA-320a对抑制神经胶质瘤U251细胞迁移、侵袭的可能的机制。实验开始前我们利用生物信息学软件进行分析对比miRNA-320a与水通道蛋白4(AQP4)之间的靶点结合关系,然后我们用miRNA-320a mimic及Ncontrol对U251细胞株进行转染,48 h后进行下一步实验。首先用q-PCR验证过表达转染情况,以及水通道蛋白4(AQP4)m RNA的表达水平,其次用划痕和Transwell检测转染后细胞株的迁移侵袭能力,最后用Western blotting测定AQP4的表达水平。生物信息分析可得miRNA-320a在AQP4 m RNA 3’UTR区域能稳定结合,实验结果显示转染mimic后,过表达组明显升高,且过表达组AQP4 m RNA的表达明显被抑制,划痕和Transwell实验提示了过表达miRNA-320a后能抑制U251细胞株的迁移侵袭能力(p<0.01)。Western blotting结果显示,过表达miRNA-320a与对照组相比能明显抑制AQP4蛋白的表达。所有研究结果提示miRNA-320a能靶向作用AQP4 m RNA 3’UTR区域,并抑制其蛋白表达,从而抑制了肿瘤细胞U251的迁移侵袭能力,为临床治疗恶性胶质瘤提供新的参考。
This study was to investigate the possible mechanism of overexpression of miRNA-320a on the migration and invasion of glioma U251 cells. Before the experiment, we used bioinformatics software to analyze the binding relationship between miRNA-320a and aquaporin 4 (AQP4). Then we transfected the U251 cell line with miRNA-320a mimic and Ncontrol, 48 h After the next experiment. The expression of AQP4 mRNA was verified by q-PCR and the migration and invasion of transfected cells were detected by scratches and Transwell. The expression of AQP4 The expression level. Bioinformatics analysis showed that miRNA-320a could stably bind to the 3’UTR region of AQP4 m RNA. The results showed that mimic transfected miRNA-320a overexpression group was significantly increased, and the expression of AQP4 m RNA in overexpression group was significantly inhibited And Transwell experiments suggested that miRNA-320a overexpression inhibited the invasion and migration of U251 cells (p <0.01). Western blotting results showed that miRNA-320a overexpression significantly inhibited the expression of AQP4 protein compared with the control group. All the results suggest that miRNA-320a can target the 3’UTR region of AQP4 m RNA and inhibit its protein expression, thus inhibiting the migration and invasion of tumor cells U251 and providing a new reference for clinical treatment of malignant gliomas.