基础医学与临床 ›› 2018, Vol. 38 ›› Issue (9): 1268-1273.

• 研究论文 • 上一篇    下一篇

SFRP5基因对小鼠肝癌细胞Hepa1-6糖代谢的影响

朱伟1,郑世霞1,柯琳秋2,汪毅2,陈琼科3,夏道涵1,王红漫1   

  1. 1. 重庆市人民医院
    2. 重庆渝中区枇巴山正街104号重庆市人民医院三院院区
    3. 重庆市人民医院三院院区
  • 收稿日期:2017-08-09 修回日期:2017-11-10 出版日期:2018-09-05 发布日期:2018-09-10
  • 通讯作者: 王红漫 E-mail:hongmanwang@126.com
  • 基金资助:
    Sfrp5/PP2A/Akt信号通路改善胰岛素抵抗的机制研究

Effects of SFRP5 on glucose metabolism in mouse hepatocellular carcinoma cell line Hepa1-6

  • Received:2017-08-09 Revised:2017-11-10 Online:2018-09-05 Published:2018-09-10
  • Contact: Hong-Man WANG E-mail:hongmanwang@126.com

摘要: 目的 探讨SFRP5过表达和表达抑制对小鼠肝癌细胞Hepa1-6糖代谢相关基因的影响。 方法 分别用空载病毒(Ad-GFP)、SFRP5过表达(Ad-SFRP5)及抑制病毒(Ad-shSFRP5)感染小鼠肝癌细胞Hepa1-6,通过real-time-PCR和Western blot测定细胞中病毒感染效率;用葡萄糖氧化酶(GOD)法测定细胞的葡萄糖摄取率(GUR);采用real-time-PCR和Western blot检测肝糖异生指标(PEPCK和G6Pase),经典胰岛素信号通路分子(InsR和AKT)的磷酸化水平。结果 在小鼠肝癌细胞He1-6中,与Ad-GFP组相比,Ad-SFRP5病毒可明显上调SFRP5基因的表达,而Ad-shSFRP5则相反。 与Ad-GFP组相比,SFRP5基因上调可提高葡萄糖摄取率,降低PEPCK和G6Pase mRNA(P<0.01)和蛋白水平(P<0.05),增强胰岛素信号通路分子InsR (P<0.01)和AKT (P<0.05)的磷酸化水平,而抑制SFRP5则作用相反。结论 SFRP5基因在Hepa1-6细胞中过表达,可改善糖代谢紊乱,降低肝细胞糖异生,增强胰岛素敏感性,改善胰岛素抵抗。

关键词: SFRP5, Hepa1-6, 糖代谢, 胰岛素抵抗

Abstract: Objective To investigate effects of over-expression/knockdown SFRP5 on glucose metabolism in hepa1-6 hepatocarcinoma cells of mice. Methods Hepa1-6 hepatocarcinoma cell lines, established using adenovinus infection tool, are divided into three groups: control group(Ad-GFP), over-expression SFRP5 group(Ad-SFRP5), and knockdown SFRP5 group(Ad-shSFRP5). Glucose uptake rate was detected by glucose oxidase method(GOD). Gene expressions at mRNA and protein levels were determined by qRT-PCR and Western blotting. Results Compared with control group, over-expression SFRP5 increased glucose uptake rate, decresed PEPCK (P<0.05) and G6Pase expression (P<0.01)and increased insulin-stimulated InsR (P<0.01), AKT (P<0.05) phosphorylation in hepa1-6 cells. Knockdown SFFRP5 group showed an opposite result. Conclusions Over-expression SFRP5 could improve glucose metabolism through decreasing hepatic gluconeogenesis and enhancing insulin signaling pathway.

Key words: SFRP5, Hepa1-6, Glucose metabolism, Insulin resistance

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