摘要
目的探讨转酮醇酶1(TKTL1)在体外培养的人类结肠癌细胞株(LoVo)生长增殖中的作用。方法构建针对TKTL1 mRNA的小干扰RNA(siRNA)质粒,将该质粒转染LoVo细胞;采用实时定量RT-PCR检测转染前后LoVo细胞转酮醇酶基因家族中转酮醇酶(TKT)、TKTL1、转酮醇酶2(TKTL2)mRNA表达水平的变化,通过流式细胞术和MTT法检测转染前后LoVo细胞周期和细胞增殖的改变。结果转染组(携带有pEGFP-C1-U6/TKTL1)、质粒对照组(携带有pEGFP-C1-U6/UC)和未转染组(未转染细胞)中TKT和TKTL2 mRNA的表达水平差异无显著性意义(P>0.05)。转染组细胞中TKTL1的表达水平与质粒对照组和未转染组相比,明显下调,且转染组LoVo细胞总转酮醇酶活性明显下降,细胞增殖明显被抑制,LoVo细胞被阻滞在G0/G1期。结论TKTL1在人类结肠癌细胞(LoVo细胞系)的生长增殖中起重要作用,TKTL1可能成为肿瘤治疗的新靶点。
Objective To investigate the effect of transketolase-like-1 （TKTL1） on proliferation of human colon cancer cell line LoVo cells in vitro. Methods siRNA plasmid against gene TKTL1 （pEGFP-C1-U6/TKTL1） was constructed and transfected into LoVo cells. The expression of TKT gene family members TKT, TKTL1 and TKTL2 mRNA was detected by realtime PCR in LoVo cells before and after transfection. The changes in proliferation and cell cycle of LoVo cells were assayed by MTT and flow cytometry before and after transfection. Results There was no significant difference in the expression level of TKT and TKTL2 among transfected cells, untransfected cells and plasmid control cells （P〉0.05）. The TKTL1 expression in transfected cells was decreased significantly as compared with that in untransfected cells and plasmid control cells. In transfected cells, the total transketolase activity was down-regulated, cell proliferation was inhibited and cells were arrested in G0/G1 phase. Conclusion TKTL1 plays an important role in the total transketolase activity and in cell proliferation of human colon cancer.
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2008年第3期347-350,共4页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
湖北省自然科学基金资助项目(No2004ABA189)
