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目的:预测和验证miR-148a-3p的靶基因。方法:利用生物信息学方法预测miR-148a-3p的靶基因;在神经胶质瘤细胞U87/U251中建立miR-148a过表达及抑制表达稳定细胞系。利用q PCR实验验证miR-148a-3p与靶基因的靶向关系;利用Western Blot实验检测靶蛋白表达水平。结果:生物信息学方法预测结果显示dynein light chain LC8-type 2(DYNLL2)为miR-148a-3p靶基因之一;q PCR实验结果显示DYNLL2在神经胶质细胞HA1800与神经胶质瘤细胞U87/U251中的表达有差异,在神经胶质瘤细胞miR-148a过表达及抑制表达的细胞中表达有差异;Western Blot实验结果显示DYNLL2的蛋白表达水平在神经胶质细胞HA1800与胶质瘤细胞U87/U251中有差异。结论:miR-148a-3p与DYNLL2有确切的靶向关系,DYNLL2在神经胶质细胞与胶质瘤细胞中均有表达,与正常胶质细胞相比,DYNLL2在神经胶质瘤细胞中表达下调,包括基因水平和蛋白水平,miR-148a-3p能够负向调控DYNLL2的表达。
Objective: To predict and validate the target gene of miR-148a-3p. Methods: The target genes of miR-148a-3p were predicted by bioinformatics methods. MiR-148a overexpression and stable cell lines were established in glioma U87 / U251 cells. The targeted relationship between miR-148a-3p and the target gene was verified by q PCR and Western Blot was used to detect the target protein expression level. Results: The results of bioinformatics method showed that dynein light chain LC8-type 2 (DYNLL2) was one of the target genes of miR-148a-3p. Q PCR results showed that DYNLL2 was expressed in glioma HA1800 and glioma U87 / U251 expression in glioma cells miR-148a overexpression and inhibition of expression in differentiated cells; Western Blot experimental results show that the expression of DYNLL2 protein levels in glioma HA1800 and glioma cells U87 / U251 in the difference. Conclusion: miR-148a-3p has an exact target relationship with DYNLL2, and DYNLL2 is expressed in glioma and glioma cells. Compared with normal glial cells, DYNLL2 is down-regulated in glioma cells , Including gene level and protein level, miR-148a-3p can negatively regulate DYNLL2 expression.