甜菜红素下调HeLa细胞线粒体丙酮酸代谢旁路蛋白编码的转录  

Betanin Down-Regulates the Protein Coding Transcripts of the Pyruvate by-Pathway in Mitochondia of HeLa Cells

作　　者：雒秋江[1,2] 包晓玮[1,2] 陈勇[1,2] 肖珊 

机构地区：[1]新疆农业大学食品科学与药学学院,乌鲁木齐830052 [2]新疆百草药用植物研究所,乌鲁木齐830052 [3]上海美吉生物医药科技有限公司,上海201203

出　　处：《中国药学杂志》2016年第16期1379-1384,共6页Chinese Pharmaceutical Journal

基　　金：新疆维吾尔自治区科技厅资助项目(2011ZD1007)

摘　　要：目的探讨甜菜红素对HeLa细胞增殖及与丙酮酸代谢和糖酵解的关系。方法在体外培养HeLa细胞,贴壁后加入0、62.5和500 mg·L^(-1)甜菜红素,48 h后收获细胞分别进行细胞计数和低温保存。以人基因组为参考,将冷冻细胞进行高通量转录组测定和计算,并利用互联网数据库,选取其糖酵解、丙酮酸代谢和柠檬酸循环部分进行分析,以研究甜菜红素对HeLa细胞增殖、糖酵解与氧化途径蛋白质编码转录本表达的影响。结果二个浓度的甜菜红素均抑制培养HeLa细胞的增殖,并且影响培养细胞的转录本reads数。HeLa细胞表达的蛋白编码转录本有74种,其中reads数大于1和reads数小于1但处理间差异显著的共40种。甜菜红素显著上调HeLa细胞糖酵解限速酶血小板型磷酸果糖激酶(ENST00000381075)和糖异生限速酶果糖-1,6-二磷酸酶1(ENST00000375326)编码蛋白转录本表达,有利于细胞代谢,可能是细胞对甜菜红素的应激反应。甜菜红素对柠檬酸循环的蛋白编码转录表达没有显著影响。然而,甜菜红素显著下调HeLa细胞线粒体乳酸脱氢酶D(ENST00000450168)(LDHD)编码蛋白转录本的表达,并且对磷酸烯醇化丙酮酸羧激酶2(线粒体)(ENST00000396973)(PEPCK)也呈下调趋势,它们均与丙酮酸代谢有关。结论甜菜红素抑制HeLa细胞的增殖与线粒体丙酮酸代谢的LDHD和PEPCK蛋白编码的转录下调有关。OBJECTIVE To study the effects of batanin on proliferation of HeLa cells and the relationships with pyruvate metabolism and glycolysis. METHODS The HeLa cells were cultured in vitro and betanin at 0, 62.5 and 500 mg · L^-1 was added after the attachment of the cells. The cells were harvested after 48 h of culture for the cell counting and storaged at low temperature. Referring to human genome, the frozen cells were carried out for the analysis and calculation of high-throughout transcriptome. Utilizing internet data base, the parts related with glyeolysis, pyruvate metabolism and citric cycle were selected and analyzed to study the effect of betanin on the protein coding transcripts of the glycolysis and citric cycle in HeLa cells. RESULTS Betanin with both of the 2 concentrations inhibited the proliferation of the cultured HeLa cells, and affected the transcript reads of the cultured ceils. There were 74 of protein coding transcripts expressed in HeLa cells, but only 40 of them had the reads 〉 1 or 〈 1 but with significant difference between treatments. Betanin significantly up-regulated the protein coding transcripts of the phosphofructokinase （platelet） （ ENST00000381075 ） and fructose-1,6-bisphosphatase 1 （ ENST00000375326 ） which were ratelimited enzymes in the glycolysis and glueoneogenesis respectively, it was benefit for cell metabolism, possibly being the stress response of the cell to betanin. However, betanin significantly down-regulated the protein coding transcript of the mitochondial lactate dehydrigenase D（LDHD） （ENST00000450168）of HeLa cell, and the mitoehondial phosphoenolpyruvate earboxykinase 2 （PEPCK） also had a tendency to being down-regulated, which all were related with pyruvate metabolism. CONCLUSION The inhibition of betanin on proliferation of HeLa cell is related with the down-regulation of the protein coding transcripts of the LDHD and PEPCK of the mitochondrial pyruvate metabolism of the cell.

关 键 词：甜菜红素 HELA 线粒体 丙酮酸 转录本 

分 类 号：R965[医药卫生—药理学]