miR⁃206 过表达靶向VEGFA 对人肝癌HepG2 细胞侵袭和迁移的抑制作用
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(1. 攀枝花市中心医院消化内科,四川攀枝花 617067; 2. 绵阳市第三人民医院检验科四川省精神卫生中心,四川绵阳 621000)

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R-33

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miR⁃206 overexpression inhibits the invasion and migration of liver cancer cell line HepG2 by targeting VEGFA
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(1. Department of Gastroenterology, Central Hospital of Panzhihua, Panzhihua 617067, China; 2. Department of Clinical Laboratory, The Third People’s Hospital of Mianyang City, Sichuan Mental Health Center, Mianyang 621000)

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    摘要:

    目的 探究miR?206 过表达对人肝癌HepG2 细胞侵袭和迁移的影响及其分子机制?方法 HepG2细胞分为四组:HepG2 空白对照组,miR?206 mimic 组,pc?VEGFA 组和miR?206 mimic + pc?VEGFA 组?qRT?PCR 检测miR?206 表达和VEGFA 的mRNA 水平?荧光素酶实验确认miR?206 与VEGFA 的靶向关系?Western blot 检测VEGFA,抗波形蛋白,N?钙粘蛋白和纤维连接蛋白的表达?Transwell 检测细胞侵袭?划痕实验分析细胞迁移?结果 转染miR?206 mimic 后HepG2 细胞中miR?206 表达显著上升,VEGFA 的mRNA 水平显著下降( P < 0. 001)?miR?206 与VEGFA 存在直接靶向关系?与对照组相比,miR?206 mimic 组VEGFA 的蛋白水平,每个视野下的侵袭细胞数目,痕愈合率显著下降( P < 0. 01)?pc?VEGFA 组VEGFA 的蛋白水平,每个视野下的侵袭细胞数目,痕愈合率显著高于对照组( P < 0. 01)?与pc?VEGFA 组相比,miR?206 mimic + pc?VEGFA 组VEGFA 的蛋白水平,每个视野下的侵袭细胞数目,痕愈合率显著降低( P < 0. 01)?miR?206 mimic 组vimentin, N?cadherin 和Fibronectin 表达低于对照组( P < 0. 01)?pc?VEGFA 组vimentin, N?cadherin 和Fibronectin 表达高于对照组( P < 0. 01)?与pc?VEGFA 组相比,miR?206 mimic + pc?VEGFA 组vimentin, N?cadherin 和Fibronectin 表达降低( P < 0. 01)?结论 miR?206过表达通过靶向VEGFA 抑制人肝癌HepG2 细胞侵袭和迁移?

    Abstract:

    Objective To explore the effect of miR?206 overexpression on the invasion and migration of the liver cancer cell line HepG2. Methods HepG2 cells were divided into four groups: HepG2 (control) group, miR?206 mimic group, pc?VEGFA group, and miR?206 mimic + pc?VEGFA group. The expression of miR?206 and level of VEGFA mRNA were analyzed by quantitative real?time reverse?transcription PCR. A luciferase experiment was used to confirm the targeting of VEGFA by miR?206. The expression of VEGFA, vimentin, N?cadherin, and fibronectin was detected by western blotting. Cell invasion was measured by a Transwell assay. Cell migration was determined by a scratch assay. Results After transfection with miR?206 mimic in HepG2 cells, the expression of miR?206 was increased with declined mRNA level of VEGFA ( P < 0. 001). VEGFA was a direct target of miR?206. Compared with control group, the protein level of VEGFA, the number of invasive cells per field and scratch healing rate in miR?206 mimic group was reduced ( P < 0. 01). The protein level of VEGFA, the number of invasive cells per field and scratch healing rate in pc?VEGFA group was higher than control group ( P < 0. 01). Compared with pc?VEGFA group, the protein level of VEGFA, the number of invasive cells per field and scratch healing rate in miR?206 mimic + pc?VEGFA group was decreased ( P < 0. 01). The expression of vimentin, N?cadherin and Fibronectin in miR?206 mimic group was lower than control group ( P < 0. 01). The expression of vimentin, N?cadherin and Fibronectin in pc?VEGFA group was higher than control group ( P < 0. 01). Compared with pc?VEGFA group, the expression of vimentin, N?cadherin and Fibronectin in miR?206 mimic + pc?VEGFA group was reduced ( P < 0. 01). Conclusions The overexpression of miR?206 inhibits the invasion and migration of liver cancer HepG2 cells by targeting VEGFA.

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王小明,余珊,赵晓姬. miR⁃206 过表达靶向VEGFA 对人肝癌HepG2 细胞侵袭和迁移的抑制作用[J].中国比较医学杂志,2018,28(8):95~100.

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  • 收稿日期:2018-03-15
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  • 在线发布日期: 2018-09-17
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