低氧诱导胶质瘤细胞上皮间质转化及差异表达的长非编码RNA的筛选

Differential expression of long non-coding RNA regulates hypoxia-induced epithelial-mesenchymal transition in glioma cells

目的探索低氧诱导条件下神经胶质瘤细胞发生上皮间质转化相关的长非编码RNA与其侵袭转移的关系。方法通过芯片检测低氧诱导前后胶质瘤细胞的长非编码RNA的表达差异。通过qRT-PCR验证芯片结果的准确性以及上皮间质转化相关基因的表达变化。通过转染敲低长非编码RNA的表达水平。通过蛋白免疫印迹检测蛋白水平的表达变化。通过侵袭迁移实验检测侵袭迁移能力的变化。结果低氧诱导神经胶质瘤细胞可发生上皮间质转化,芯片检测发现多个差异表达的长非编码RNA,长非编码RNA HOTTIP在低氧诱导后表达升高最为明显(P<0.01),降低其表达水平后可逆转其上皮间质转化的过程,并显著降低侵袭迁移能力(P<0.01)。结论长非编码RNA HOTTIP的表达升高可能与低氧诱导所发生的上皮间质转化以及增强的侵袭迁移能力密切相关。

Objective To investigate the correlation between hypoxia-induced epithelial-mesenchymal transition associated long non-coding RNA in glioma cells and its capacities of invasion and metastasis. Methods The expression of long non-coding RNA in glioma cells was detected by microarray. The accuracy of the results of microarray and the expression of epithelial-mesenchymal transition related genes were verified by qRT-PCR. The expression of proteins was detected by Western blotting. The invasion and migration capacities were assessed by invasion and migration assays. Results Hypoxia induced epithelial-mesenchymal transition of glioma cells. A number of differentially expressed long non-coding RNAs were found and long non-coding RNA HOTTIP was found up-regulated after hypoxia induction(P<0.01). Reducing its expression level reversed the process of epithelial mesenchymal transition, and significantly reduced the invasion and migration capacities(P<0.01). Conclusion The increased expression of long non-coding RNA HOTTIP may be closely related to hypoxia-induced epithelial-mesenchymal transition and enhanced invasion and migration.