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目的:通过RNA干涉下调CD44在脑胶质瘤细胞A172中的表达,观察CD44下调后对胶质瘤细胞增殖、迁移和侵袭的影响。方法:根据CD44基因序列设计并合成的siR-NA片段(CD44-siRNA)转染A172细胞,利用荧光实时定量qRT-PCR检测细胞中CD44基因mRNA水平的变化;Westernblot检测CD44蛋白水平的变化;MTT法检测A172细胞增殖;流式细胞术(FCM)检测A172细胞周期与凋亡。结果:CD44-siRNA下调了A172细胞中CD44 mRNA水平与蛋白水平的表达,A172细胞的增殖能力明显降低,并将A172细胞阻滞于G0/G1期,A172细胞的凋亡数量明显增加,尤其是晚期凋亡细胞数。结论:CD44-siRNA能够下调CD44的表达,有效抑制A172脑胶质瘤细胞的增殖,并促进其凋亡,为以CD44为靶点的脑胶质瘤基因治疗提供理论依据。
OBJECTIVE: To investigate the effect of CD44 on the proliferation, migration and invasion of glioma cells by down-regulating the expression of CD44 in A172 glioma cells by RNA interference. Methods: A172 cells were transfected with siR-NA fragment (CD44-siRNA) designed and synthesized based on the sequence of CD44 gene. Fluorescence quantitative real-time qRT-PCR was used to detect the mRNA expression of CD44. Western blot was used to detect the level of CD44 protein. MTT A172 cell proliferation assay; flow cytometry (FCM) A172 cell cycle and apoptosis. Results: The expression of CD44 mRNA and protein in A172 cells was down-regulated by CD44-siRNA, the proliferation of A172 cells was significantly reduced, and A172 cells were arrested in G0 / G1 phase. The number of apoptosis in A172 cells was significantly increased, especially Late apoptotic cells. Conclusion: CD44-siRNA can downregulate the expression of CD44, effectively inhibit the proliferation of A172 glioma cells and promote its apoptosis, providing a theoretical basis for the gene therapy of glioma targeting CD44.