摘要
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 explore the effects and regulation mechanism of miRNA-376c-3p on proliferation,migration and apoptosis of human gastrointestinal stromal tumor(GIST)cell line GISTT1.Methods:The expression of miRNA-376c-3p in tumor tissues and adjacent normal tissues of13 gastric stromal tumor patients was detected by using Real-time quantitative PCR.GIST-T1 cells were allocated into miR-376c-3p mimic and miR-376c-3p inhibitor transfected group,as well as negative control and blank control group.MiR-376c-3p expression in each group was determined by qRT-PCR.Cell proliferation was evaluated by using the CCK8 assay and apoptosis was accessed by flow cytometry.Tumor cell migration was measured by Transwell chamber assays.Furthmore,the effects of miR-376c-3p on CASP-7(Caspase-7)expression were examined by qRTPCR and Western Blot analysis.Results:As shown by qRT-PCR,compared with that in the gastric stromal tumor adjacent tissue,the expression of miR-376c-3p in the tumor tissue was significantly decreased(P<0.01).Meanwhile,miR-376c-3p mRNA expression in GIST-T1 cells transfected with miR-376c-3p mimic or inhibitor was significantly higher or lower than the negative control and blank control group(P<0.01).The proliferation and migration of GIST-T1 cells were dramatically suppressed by up-regulation of miR-376c-3p(P<0.01)while cell apoptotic rates were obviously increased(P<0.05).In addition,CASP-7mRNA and protein expression were significantly increased after miR-376c-3p mimic transfectionin contrast with down-regulation of CASP-7mRNA and protein by inhibition of miR-376c-3p(P<0.05).Conclusion:MiR-376c-3p could inhibit cell proliferation,migration and induce apoptosis of GIST-T1 cells via regulating expression of CASP-7.
出处
《武汉大学学报（医学版）》
CAS
2017年第2期231-236,共6页
Medical Journal of Wuhan University
基金
湖北省卫计委资助项目(编号:WJ2015MB030)
