miR-124通过靶向Capn4抑制Wnt/β-catenin信号通路调节胃肠道间质瘤(GIST-T1)细胞生物学功能

miR-124 regulates biological activity of human gastrointestinal stromal tumor GIST-T1 cell line by targeting Capn4 and inhibiting Wnt/β-catenin signaling

目的:探讨miR-124对人胃肠道间质瘤GIST-T1细胞增殖、迁移和凋亡的影响以及其可能的调控机制。方法:qRT-PCR检测30例胃肠道间质瘤患者的肿瘤和癌旁对照组织中的miR-124表达;利用脂质体试剂Lipofectamine 2000将miR-124 mimic、miR-124 inhibitor及其对应的阴性对照分别转入GIST-T1细胞,并且设置空白对照;qRT-PCR检测细胞中miR-124的表达情况;CCK-8检测细胞增殖;Transwell检测细胞侵袭能力;流式细胞术检测细胞模拟物或抑制剂处理后的细胞凋亡情况;Western blot检测Capn4蛋白表达情况。结果:miR-124在肿瘤组织中显著低于癌旁对照组织;过表达miR-124后,CCK-8实验及Transwell实验结果显示GIST-T1细胞增殖、迁移能力和侵袭能力均显著下降;流式细胞术结果显示GIST-T1细胞晚期凋亡降低,早期凋亡增加;Western blot结果显示过表达miR-124后,GIST-T1细胞中Capn4蛋白表达明显降低,当抑制miR-124后,GIST-T1细胞中Capn4蛋白表达明显上升;抑制miR-124后GIST-T1细胞中β-catenin蛋白表达增加。结论:miR-124通过靶向Capn4抑制GIST-T1细胞增殖、迁移和侵袭,并促进细胞凋亡,与其抑制Wn/tβ-catenin信号通路有关。

Objective:To explore the effects and regulation mechanism of miR-124 on proliferation,migration,invasion and apoptosis of human gastrointestinal stromal tumor(GIST)cell line GIST-T1.Methods:The expression of miR-124 in tumor tissues and adjacent normal tissues of 30 patients was detected by using qRT-PCR.GIST-T1 cells were allocated into miR-124 mimic and inhibitor transfected group,as well as negative control and blank control group by liposome reagent Lipofectamine 2000.miR-124 expression was detected by qRT-PCR in cells.Cell proliferation was determined by CCK-8 assay.Cell migration and invasion was detected by Transwell assay.Cell apoptosis was evaluated by flow cytometry,and Western blot was used to detect Capn4 protein expression.Results:miR-124 expression was significantly lower in tumor tissues than in the gastrointestinal stromal tumor adjancent tissues.After overexpression of miR-124,the results showed the proliferation,migration and invasion of GIST-T1 cells were significantly decreased while cell apoptotic rates were dramatically increased.Western blot showed that Capn4 protein expression were significantly decreased after miR-124 mimics transfection contrast with down-regulation of Capn4 protein expression by inhibition of miR-124 in cells.Theβ-catenin protein expression in GIST-T1 cells was increased after inhibition of miR-124.Conclusion:miR-124 inhibits GIST-T1 cell proliferation,migration and invasion,promotes cell apoptosis by targeting Capn4,which relative with inhibiting Wn/tβ-catenin signaling pathway.