miR-873-5p通过靶向自噬相关基因Beclin1抑制肺癌细胞进展和抗血管生成的机制  

Mechanism of miR-873-5p inhibiting progression and anti-angiogenesis of lung cancer cells by targeting autophagy-related gene Beclin1

作　　者：王永杰 方小丽 刘仕强 王杰 陈旭 汪华 何文武 WANG Yong-jie;FANG Xiao-li;LIU Shi-qiang;WANG Jie;CHEN Xu;WANG Hua;HE Wen-wu(Department of Thoracic surgery,Nanchong Central Hospital of Sichuan Province,Nanchong,Sichuan Province 1637000,China;Department of Cardiology Medicine,Nanchong Central Hospital of Sichuan Province,Nanchong,Sichuan Province 1637000,China;Department of Thoracic Surgery,Sichuan Cancer Hospital,Chengdu,Sichuan Province 3610000)

机构地区：[1]四川省南充市中心医院胸心外科,四川南充1637000 [2]四川省南充市中心医院心内科,四川南充2637000 [3]四川省肿瘤医院胸外科,四川成都3610000

出　　处：《解剖学研究》2023年第1期27-33,共7页Anatomy Research

摘　　要：目的 探讨微小RNA(miRNA)-873-5p通过靶向自噬相关基因Beclin1抑制肺腺细胞进展和抗血管生成的机制研究。方法 通过双荧光素酶报告验证miR-873-5p靶向Beclin1。将人肺癌细胞系A549随机分为4组:对照组、Mimic组、Mimic+Beclin1组和Beclin1组。通过质粒转染技术过表达miR-873-5p以及Beclin1。实时荧光定量PCR(q PCR)和Western blot检测RNA或蛋白的表达水平。分别通过CCK-8、克隆形成实验和流式细胞术检测细胞活力、增殖和凋亡,细胞划痕实验、Transwell实验和管生成实验检测各组的细胞迁移、侵袭和血管生成能力。结果 MiR-873-5p直接靶向Beclin1。过表达miR-873-5p抑制Beclin1 mRNA和蛋白的表达水平,过表达Beclin1显著逆转miR-873-5p对Beclin1的抑制作用。Mimic组的细胞活力和细胞增殖能力显著低于对照组,凋亡率高于对照组(P<0.05),Beclin1组的细胞活力和细胞增殖能力显著高于对照组,细胞凋亡率低于对照组(P<0.05),并且Mimic+Beclin1组的细胞活力和细胞增殖能力显著高于Mimic组,细胞凋亡率低于mimc组(P<0.05)。Mimic组的细胞迁移和侵袭能力显著低于对照组(P<0.05),Beclin1组的细胞迁移和侵袭能力显著高于对照组(P<0.05),并且Mimic+Beclin1组的细胞迁移和侵袭能力显著高于Mimic组(P<0.05);Mimic组的血管生成能力显著低于对照组(P<0.05),Beclin1组的血管生成能力显著高于对照组(P<0.05),并且Mimic+Beclin1组的血管生成能力显著高于Mimic组(P<0.05)。结论 MiR-837-5p可通过靶向抑制Beclin1的水平抑制肺癌细胞的生长、侵袭和血管生成,从而发挥抑制肺癌转移的作用。Objective To explore the mechanism of microRNA(miRNA)-873-5p inhibition of lung gland cell progression and anti-angiogenesis by targeting autophagy-related gene Beclin1. Methods The miR-873-5p was targeted to Beclin1 by dual luciferase reporter. Human lung cancer cell line A549 was divided into four groups:control group,Mimic group,Mimic+Beclin1 group and Beclin1 group. miR-873-5p and Beclin1 were overexpressed by plasmid transfection techniques. Quantitative Real-time PCR(q PCR)and Western blot were used to detect the expression level of Ribonucleic Acid(RNA)or protein. Cell viability,proliferation and apoptosis were detected by cell counting kit-8(CCK-8)assay,colony formation assay and flow cytometry,respectively. Cell scratch assay,Transwell assay and tube generation assay were used to detect cell migration,invasion and angiogenesis in each group.Results MiR-873-5p directly targets Beclin1. Overexpression of miR-873-5p inhibits the expression levels of Beclin1 mRNA and protein. Overexpression of Beclin1 significantly reversed the inhibitory effect of miR-873-5p on Beclin1. The cell viability and cell proliferation ability of the Mimic group were significantly lower than those of the control group,and the apoptotic rate was higher than that of the control group(P<0.05). The cell viability and cell proliferation ability of Beclin1 group were significantly higher than that of the control group,and the apoptosis rate was lower than that of the control group(P<0.05). The cell viability and cell proliferation ability of Mimic+Beclin1group were significantly higher than that of Mimic group,and the apoptosis rate was lower than that of mimc group(P<0.05). The cell migration and invasion ability of the Mimic group was significantly lower than that of the control group(P<0.05). The cell migration and invasion ability of Beclin1 group was significantly higher than that of the control group(P<0.05). The cell migration and invasion ability of Mimic+Beclin1 group was significantly higher than that of Mimic group(P<0.05). Th

关 键 词：肺癌 非小细胞肺癌 微小RNA 自噬基因BECLIN1 自噬 血管生成 

分 类 号：R734.2[医药卫生—肿瘤]