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目的:探讨miRNA-376c-3p(miR-376c-3p)对人胃肠道间质瘤GIST-T1细胞迁移、增殖和凋亡的影响及其可能的调控机制。方法:qRT-PCR法检测13例胃间质瘤患者肿瘤和癌旁对照组织中miR-376c-3p表达;将miR-376c-3p类似物(mimic)、抑制物(inhibitor)和各自阴性对照分别转染GIST-T1细胞,另设空白对照组;qRT-PCR检测转染细胞miR-376c-3p表达;然后用CCK8检测细胞增殖能力,流式细胞术检测细胞凋亡,Transwell小室检测细胞迁移能力;通过Western Blot及qRT-PCR法检测miR-376c-3p对CASP-7(Caspase-7)表达量的影响。结果:在胃间质瘤患者癌组织中,miR-376c-3p表达水平显著降低,为癌旁对照组织的58.7%(P<0.01);qRT-PCR显示:转染miR-376c-3p mimic后,GIST-T1细胞miR-376c-3p表达明显上调(P<0.01),而转染miR-376c-3p inhibitor后表达受抑制(P<0.01);miR-376c-3p mimic转染组细胞增殖活性、迁移能力明显抑制(P<0.01),凋亡率增强(P<0.05);另外miR-376c-3p过表达后CASP-7mRNA和蛋白水平出现上调,相反地,miR-376c-3p抑制却导致CASP-7mRNA和蛋白水平下调(P<0.05)。结论:在人胃肠道间质瘤GIST-T1细胞中miR-376c-3p可以间接调控CASP-7的表达,抑制细胞增殖、迁移能力,促进细胞凋亡。
Objective: To investigate the effects of miRNA-376c-3p (miR-376c-3p) on the migration, proliferation and apoptosis of human gastrointestinal stromal tumors (GIST-T1) cells and its possible regulatory mechanism. Methods: qRT-PCR was used to detect the expression of miR-376c-3p in 13 cases of gastric stromal tumors and adjacent non-cancerous tissues. The miR-376c-3p analogs (mimic), inhibitor and their negative control The expression of miR-376c-3p in transfected cells was detected by qRT-PCR. The proliferation of transfected cells was detected by CCK8. The apoptosis was detected by flow cytometry. The transwell chamber was used to detect the cell migration The effect of miR-376c-3p on Caspase-7 expression was detected by Western Blot and qRT-PCR. Results: The expression of miR-376c-3p was significantly decreased in cancer tissues of patients with gastric stromal tumors (58.7%, P <0.01). QRT-PCR showed that miR-376c-3p mimic The expression of miR-376c-3p in GIST-T1 cells was significantly up-regulated (P <0.01), while the expression of miR-376c-3p inhibitor was inhibited after transfected with miR-376c-3p mimic (P <0.01). The apoptotic rate of miR-376c-3p was up-regulated after miR-376c-3p overexpression. On the contrary, the inhibition of miR-376c-3p resulted in CASP-7 mRNA and protein levels were down-regulated (P <0.05). Conclusion: miR-376c-3p indirectly regulates the expression of CASP-7 in human gastrointestinal stromal tumors GIST-T1 cells, inhibits cell proliferation and migration, and promotes apoptosis.