醛缩酶A基因促进肝癌细胞增殖  

Aldolase A gene affects the growth of hepatocellular carcinoma cells

作　　者：曾诚[1,2] 刘文娟[1,2] 李小强[1,2] 张晓文[1,2,3] 谢艳华[1,2] 曹蔚[1,2] 

机构地区：[1]第四军医大学药学院天然药物学教研室,西安710032 [2]第四军医大学药学院药理学教研室,西安710032 [3]陕西省商洛学院生物医药与食品工程学院,商洛726000

出　　处：《国际药学研究杂志》2016年第6期1088-1092,共5页Journal of International Pharmaceutical Research

基　　金：国家自然科学基金资助项目(81473329);陕西省社会发展科技攻关项目(2016SF-284)

摘　　要：目的初步探索醛缩酶A基因敲除与过表达对肿瘤细胞生长的影响,确定无氧代谢过程中醛缩酶与肿瘤生长代谢之间的关系。方法通过表达载体质粒构建与慢病毒感染构建稳定表达的醛缩酶A过表达与基因敲除细胞,在基因和蛋白水平验证重组细胞,观察细胞增殖速度改变。结果实时定量PCR结果显示,过表达细胞系中目的基因的表达约为正常细胞的2倍;根据醛缩酶A RNA序列设计5个干扰靶点,其中醛缩酶A4对于目的基因的干扰效果最为显著,重组后的细胞醛缩酶A mRNA表达水平为正常细胞的1/50。与正常对照组相比,醛缩酶A过表达细胞在48 h增殖率增加40%,增殖率与对照组相比有显著性差异(P<0.05);醛缩酶A RNA干扰表达细胞在24 h增殖率降低86%,与对照组相比有显著性差异(P<0.05)。结论醛缩酶A的表达差异对肝癌肿瘤细胞生长有明显影响。Objective To preliminarily explore the effects of gene knock-out and overexpression of aldolase A on tumor cell proliferation,and then confirm the relationship between aldolase A and tumor growth during the process of anaerobic metabolism.Methods Through vector plasmid building and lentiviral infection,the cell lines stably overexpressing aldolase A and its gene knock-out cell lines were constructed respectively. The restructured cell lines were validated on gene and protein levels. Then,the changes of cell proliferation ratio were observed. Results Real time PCR results showed that aldolase A gene expression of the overexpressing cell line was about 3 times that of the normal cells. Among the five interfering targets of aldolase A designed according to aldolase A RNA sequences,aldolase A4 had more obvious interference effect on the target gene expression,the aldolase A m RNA level of which was about 1/50 of the normal cells. Compared with normal control group,the proliferation rate of aldolase A gene overexpression significantly increased by 40% in 48 h（P〈0.05）,while the proliferation rate of aldolase A gene knock out cells significantly decreased about 86% in 24 h（P〈0.05）. Conclusion Aldolase A expressions have obvious effects on hepatocellular carcinoma cells.

关 键 词：醛缩酶 基因敲除 过表达 肝癌细胞 

分 类 号：R735.7[医药卫生—肿瘤]