摘要
结肠癌是常见的消化道恶性肿瘤。目前,结肠癌早期诊断和靶向治疗仍存在不足,使其发病率占所有恶性肿瘤第3位,死亡率位于第5位,给人类的健康带来巨大的威胁。及早发现结肠癌治疗的新靶点对其预后至关重要。已有研究揭示,微RNA-149(miR-149)在结肠癌患者中低表达与结肠癌的不良预后呈正相关,但其在结肠癌发生发展过程中的作用机制有待阐明。基于此,本研究首先通过MTT和集落形成结果发现,miR-149对结肠癌HT-29细胞的增殖具有显著的抑制作用(P<0.05)。线粒体荧光标记及Western免疫印迹结果表明,miR-149通过促进线粒体融合蛋白2 (mitofusin2,MFN2)的表达,抑制线粒体动力相关蛋白1(dynamin-related protein1,DRP1)和线粒体分裂蛋白1(mitochondrial fission protein1,FIS1)的表达,进而诱导了结肠癌HT-29细胞线粒体的融合(P<0.05)。此外,通过细胞线粒体膜电位检测和蛋白质免疫印迹分析线粒体的功能状态结果显示,miR-149可以促使线粒体膜电位水平降低(P<0.05)。进一步通过流式细胞术和蛋白质免疫印迹检测结肠癌HT-29细胞的凋亡情况。结果显示,miR-149诱导了结肠癌HT-29细胞的凋亡(P<0.001)。综上,本研究从调控线粒体结构的角度出发,阐明了miR-149通过促进线粒体融合发挥抗结肠癌HT-29细胞的分子机制,为miR-149进一步成为肿瘤标记物或治疗靶点奠定实验基础和理论依据。
Colon cancer is a common malignant tumor of the digestive tract.It ranks third in the incidence of all malignant tumors and the fifth in the mortality rate,which brings huge threat to human health.At present,the early diagnosis and targeted therapy of colon cancer are still insufficient.Early detection of new targets for colon cancer therapy is critical to its prognosis.Previous studies have revealed that low expression of miR-149(microRNA-149) in patients with colon cancer is positively correlated with poor prognosis,but its mechanism in the development and progression of colon cancer remains to be clarified.In this study,MTT assays and colony formation assays showed that miR-149 inhibited the proliferation ability of colon cancer HT-29 cells(P<0.05).Mitochondrial fluorescence labeling and Western blot results showed that miR-149 induced mitochondrial fusion in colon cancer HT-29 cell by promoting the expression of mitochondrial fusion protein2(mitofusin2,MFN2) and inhibiting the expression of mitochondrial dynamin-related proteins(dynamin-related protein1,DRP1) and mitochondrial fission proteins(mitochondrial fission protein1,FIS1)(P<0.05).In addition,the functional status of mitochondria was analyzed by mitochondrial membrane potential detection and Western blot.The results showed that miR-149 promoted a decrease of mitochondrial membrane potential(P<0.05).The apoptosis of colon cancer HT-29 cell was further detected by flow cytometry and Western blot.The results showed that miR-149 induced the apoptosis of colon cancer HT-29 cells(P<0.001).In conclusion,from the perspective of regulating mitochondrial structure,this study clarified the anti-cancer effect of miR-149 by promoting mitochondrial fusion,which lays an experimental and theoretical basis that miR-149 may become a tumor marker or therapeutic target in colon cancer.
作者
史江颖
毕彩
纪晓丹
单树花
李卓玉
SHI Jiang-Ying;BI Cai;JI Xiao-Dan;SHAN Shu-Hua;LI Zhuo-Yu(Institute of Biotechnology,Key Laboratory of Chemical Biology and Molecular Engineering of National Ministry of Education,Shanxi University,Taiyuan 030006,China)
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2023年第1期121-129,共9页
Chinese Journal of Biochemistry and Molecular Biology
基金
国家自然科学基金(No.81803238,32072220)
山西省自然科学基金面上项目(No.202103021224011)。
关键词
微RNA-149
线粒体融合
结肠癌
凋亡
micro RNA-149(miR-149)
mitochondrial fusion
colon cancer
apoptosis
作者简介
通讯作者:李卓玉,Tel:0351-7018268,E-mail:lzy@sxu.edu.cn。
