摘要
目的通过检测miR-130a的表达情况及miR-130a与肿瘤抑制因子(CYLD)的靶向关系,探讨miR-130a是否可通过调控CYLD对结直肠癌SW480细胞生物学行为产生影响。方法将结直肠癌SW480细胞设置为对照组、阴性转染组、miR-130a inhibitor组,采用qRT-PCR检测各组miR-130a的表达;MTT法、Transwell法、流式细胞术分别检测各组SW480细胞增殖、凋亡、侵袭、迁移情况;双荧光素酶报告基因检测miR-130a与CYLD靶向关系;Western blot检测各组细胞CYLD、增殖、凋亡、侵袭迁移相关蛋白表达情况。结果与对照组相比,miR-130a inhibitor组的miR-130a表达水平显著降低(P<0.05)。与对照组相比,miR-130a inhibitor组SW480细胞凋亡率显著增加,而存活率、侵袭迁移数显著降低(P<0.05)。与对照组相比,miR-130a inhibitor组c-Myc、CyclinD1、MMP7、Bcl-2表达水平显著降低,Bax、Caspase-3、CYLD表达水平显著增加(P<0.05)。靶基因预测结果显示,CYLD是miR-130a的潜在目标基因,与pmirGLO-CYLD-wt+miR-NC组相比,pmirGLO-CYLD-wt+miR-130a mimics组荧光素酶活性显著降低。结论miR-130a表达沉默可能通过促进CYLD表达抑制SW480细胞的增殖、侵袭、迁移,促进细胞凋亡。
Objective To detect the expression of miR-130a and the targeting relationship between miR-130a and cylindromatosis(CYLD),and to investigate whether miR-130a can affect the biological behavior of colorectal cancer SW480 cells by regulating CYLD.Methods SW480 cells were divided into control group,negative transfection group and miR-130a inhibitor group,the expression of miR-130a was detected by qRT-PCR;the proliferation,apoptosis,invasion and migration of SW480 cells were detected by MTT method,Transwell method and flow cytometry;double luciferase reporter genes were used to detect double luciferase reporter gene;the expression of CYLD,proliferation,apoptosis,invasion and migration related proteins were detected by Western blot.Results Compared with that in the control group,the expression level of miR-130a in the miR-130a inhibitor group was significantly lower(P<0.05).Compared with those in the control group,the apoptosis rate of SW480 cells in miR-130a inhibitor group was significantly higher,while the survival rate and invasion migration number were significantly lower(P<0.05).Compared with those in the control group,the expression levels of c-Myc,CyclinD1,MMP7 and Bcl-2 in miR-130a inhibitor group were significantly lower,and the expression levels of Bax,Caspase-3 and CYLD were significantly higher(P<0.05).The results of target gene prediction showed that CYLD was a potential target gene of miR-130a,and compared with that of pmirGLO-CYLD-wt+miR-NC group,the luciferase activity of pmirGLO-CYLD-wt+miR-130a mimics group was significantly lower.Conclusion MiR-130a silencing may inhibit the proliferation,invasion and migration of SW480 cells and promote the apoptosis of SW480 cells by promoting the expression of CYLD.
作者
李子银
习隽丽
吕志华
彭波
LI Zi-yin;XI Jun-li;LV Zhi-hua;PENG Bo(Department of Gastroenterology,the Third Hospital of Wuhan,Hubei,430000;Department of Gastroenterology,Hubei Provincial People′s Hospital,Wuhan,Hubei,430060)
出处
《现代消化及介入诊疗》
2021年第4期454-459,共6页
Modern Interventional Diagnosis and Treatment in Gastroenterology
基金
武汉市卫生和计划生育委员科研项目(WX16D10)。
