摘要
目的研究miR-506在结肠癌中的表达及其对结肠癌细胞自噬的影响。方法收集74例结肠癌患者的癌组织和癌旁组织标本,采用RT-PCR检测miR-506和Beclin1的mRNA表达水平,并分析二者的相关性。培养结肠癌细胞系HCT116、SW48、COLO225细胞,分别转染miR-506 mimic、miR-506 inhibitor或miR-control,采用RT-PCR检测Beclin1的mRNA表达水平,并用Target scan分析Beclin1是否可能与miR-506结合。培养COLO225细胞,分别转染miR-506 mimic、miR-506 inhibitor或miR-control,采用Western blot检测p62、Beclin1和LC3BⅠ/Ⅱ的蛋白表达水平。结果结肠癌患者癌组织标本miR-506、Beclin1的mRNA表达水平显著高于癌旁组织标本[(0.607±0.092)比(0.251±0.089)、(0.546±0.121)比(0.266±0.156),t=23.995、12.203,均P<0.001],而且二者的表达呈显著正相关(r=0.786、P<0.001)。在转染miR-506 mimic后,HCT116、SW48、COLO225细胞中Beclin1的表达均显著升高(P<0.05),在转染miR-506 inhibitor后,HCT116、SW48、COLO225细胞中Beclin1的表达均显著降低(P<0.05),Target scan分析表明,Beclin1上存在可与miR-506相互结合的序列。在转染miR-506 mimic后,COLO225细胞中p62、Beclin1的表达显著升高(P<0.05),LC3BⅠ向LC3BⅡ转化增加;在转染miR-506 inhibitor后,COLO225细胞中p62、Beclin1的表达显著降低(P<0.05),LC3BⅡ向LC3BⅠ转化增加。结论miR-506可促进结肠癌中Beclin1的表达并可促进结肠癌细胞自噬。
Objective To study the expression of miR-506 and its effect on autophagy in colon cancer.Methods The expression of miR-506 and Beclin1 was determined by RT-PCR in cancerous and paracancerous tissues from 74 patients with colon cancer.The correlation between miR-506 and Beclin1 were analyzed.Colon cancer cell lines HCT116,SW48 and COLO225 were cultured and transfected with miR-506 mimic,miR-506 inhibitor or miR-control.Target scan was performed to analyze whether Beclin1 bound to miR-506.Western blot was carried out to determine the protein expression of p62,Beclin1 and LC3BⅠ/Ⅱ.Results The expression of miR-506 and Beclin1 in cancerous tissues was higher than that in paracancerous tissues[(0.607±0.092)vs(0.251±0.089)and(0.546±0.121)vs(0.266±0.156),respectively;t=23.995 and 12.203,respectively;P<0.001].There was a positive correlation between miR-506 and Beclin1 expression(r=0.786,P<0.001).The expression of Beclin1 increased after transfection with miR-506 mimic and decreased after transfection with miR-506 inhibitor in HCT116,SW48 and COLO225 cells(P<0.05).Target scan showed that Beclin1 might bind to miR-506.Transfection with miR-506 mimic increased the expression of p62 and Beclin1 and promoted the conversion of LC3BⅠto LC3BⅡin COLO225 cells(P<0.05).However,transfection with miR-506 inhibitor reduced the expression of p62 and Beclin1 and promoted the conversion of LC3BⅡto LC3BⅠ(P<0.05).Conclusion miR-506 up-regulates Beclin1 expression and promotes autophagy in colon cancer.
作者
赵德余
王剑峰
郭旭
ZHAO De-yu;WANG Jian-feng;GUO Xu(Department of Anorectal Diseases,Dalian Central Hospital,Dalian 116023,China)
出处
《南昌大学学报（医学版）》
CAS
2020年第4期70-73,80,共5页
Journal of Nanchang University：Medical Sciences
