Mfn2基因沉默对HepG_2细胞葡萄糖代谢和胰岛素信号通路的影响  

Effects of Mfn2 Gene Silencing on Glucose Metabolism and Insulin Signaling Pathway in HepG_2 Cells

作　　者：陈小琳[1] 雷幼蓉[1] 

机构地区：[1]武汉大学人民医院内分泌科,430060

出　　处：《医学研究杂志》2012年第9期71-74,共4页Journal of Medical Research

基　　金：湖北省自然科学基金资助项目(2008CDB128)

摘　　要：目的研究线粒体融合素基因-2(mitofusin-2,Mfn2)沉默对人肝癌细胞株(HepG2)葡萄糖代谢和胰岛素信号通路分子丝/苏氨酸蛋白激酶(Akt1)的影响。方法构建Mfn2短发夹双链RNA(Mfn2shRNA)和阴性对照短发夹双链RNA(HK),将HepG2细胞株分为空白对照组(NC)、阴性对照组(HK)、Mfn2shRNA质粒转染组(Mfn2)。HK和Mfn2组细胞应用lipo-fectamine2000转染HepG2细胞株;采用葡萄糖氧化酶的方法和氚标记葡萄糖(3-3H-Glucose)放射性核素示踪法检测细胞葡萄糖消耗量和摄取率;Western blotting检测Mfn2和Akt1蛋白表达。结果与HK组比较,Mfn2组细胞Mfn2蛋白表达明显下降(0.23±0.05 vs 0.51±0.14,P=0.034),Akt1表达差异无统计学意义(0.13±0.00 vs 0.14±0.01,P=0.055);Mfn2组细胞葡萄糖消耗量明显下降(8.72±0.62 vs 9.75±0.35,P=0.001),葡萄糖摄取率亦显著下降(7.94±0.04 vs 9.64±0.13,P=0.002)。结论抑制Mfn2基因表达导致HepG2细胞葡萄糖代谢下降;Mfn2表达对葡萄糖代谢的影响是否通过PI3K/Akt信号通路介导还需要进一步研究。Objective To study the effects of Mitofusin - 2 gene （Mfn2） silencing on glycometabolism and Aktl （protein Ser/Thr ki- nase B, one molecule in insulin signaling pathway） expression in HepG2 cells. Methods Mfn2 short hairpin RNA （shRNA） and nega- tive control green fluorescent protein （GFP） - expressed plasmid vectors were constructed. HepG2 cell lines were randomly divided into normal control group （NC） , transfection group （Mfn2） and negative Control group （HK）. HepG2 cells were transfected using shRNA with lipofectamine 2000. The consumption of glucose was tested by glucose oxidase method and D - （3 - ^3H） glucose was used to measure the rate of glucose uptake. Mfn2 and Aktl expression were assessed by Western blotting. Results The expression of Mfn2 in Mfn2 group was significantly decreased than that of HK group（0.23 ± 0. 05 vs 0. 51 ± 0.14, P = 0. 034）. However, the difference had no statistical significant difference in the expression of Aktl protein between two groups （0.13 ± 0. 0 vs 0. 14 ± 0. 01 ,P = 0. 055 ）. Compared with the HK cells, The consumption of glucose in Mfn2 cells were markedly lower （8.72 ±0.62 vs 9.75 ±0.35, P =0. 001 ）. The rate of glucose uptake was also significantly decreased in Mfn2 cells, compared with H K cells （7.94 ± 0.04 vs 9.64 ± 0. 13,P = 0. 002 ）. Conclusion The inhibited expression of Mfn2 induces glycometabolic decrease. However, whether the PI3K/Akt signal pathway is involved in the effects of the expression of Mfn2 on glycometabolism remain to be studied.

关 键 词：线粒体融合素基因-2 HEPG2细胞 葡萄糖代谢 丝/苏氨酸蛋白激酶 

分 类 号：R735.7[医药卫生—肿瘤]