The TKTL1 gene influences total transketolase activity and cell proliferation in human colon cancer LoVo cells
The TKTL1 gene influences total transketolase activity and cell proliferation in human colon cancer LoVo cells
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DOI：
10.1097/cad.0b013e328013d99e
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发表时间：
2007-04-01
影响因子：
2.3
通讯作者：
Huang, Jin-Song
中科院分区：
文献类型：
--
作者：
Hu, Li-Hua;Yang, Ju-Hong;Huang, Jin-Song
构建了一个携带DNA的质粒，该质粒将被转录成针对转酮醇酶样-1 mRNA的小干扰RNA，并转染到人结肠癌细胞系中。用实时荧光定量聚合酶链反应检测转酮醇酶基因家族在人结肠癌细胞系中的mRNA表达。采用流式细胞术和3-（4，5-二甲基噻唑-2基）-2，5-二苯基溴化四氮唑检测抗转酮醇酶样1小干扰RNA对人结肠癌细胞系细胞增殖和细胞周期的影响。与对照载体转染的细胞和未转染的细胞相比，转酮酶样1基因在转染小干扰RNA转酮酶样1构建体的人结肠癌细胞系中显著下调。此外，抗转酮醇酶样-1小干扰RNA构建体显著降低了转染的人结肠癌细胞系细胞中的转酮醇酶水平，使其停滞在G（0）/G（1）期，并基本上抑制了细胞增殖。转酮醇酶和转酮醇酶样2基因在转染的人结肠癌细胞系中的表达与对照组相比差异无统计学意义（P > 0.05）。我们的数据表明，转酮醇酶样1基因在总转酮醇酶活性和人结肠癌细胞增殖中起着重要作用。转酮醇酶样-1可能作为一个新的抗癌治疗的目标。
A plasmid carrying DNA to be transcribed into a small interfering RNA against transketolase-like-1 mRNA was constructed and transfected into a human colon cancer cell line. The mRNA expression of transketolase gene family in the human colon cell line was determined by real-time polymerase chain reaction. The effect of anti-transketolase-like-1 small interfering RNA on cell proliferation and cell cycle in the human colon cancer cell line cells was detected by flow cytometry and 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl tetrazolium bromide. The transketolase-like-1 gene was significantly downregulated in human colon cancer cell line cells transfected with small interfering RNA transketolase-like-1 constructs compared with the cells transfected with control vector and the cells without transfection. In addition, the anti-transketolase-like-1 small interfering RNA construct significantly decreased the level of transketolase in the transfected human colon cancer cell line cells, arrested them in G(0)/G(1) phase and substantially inhibited cell proliferation. No significant difference was found in the other two genes (transketolase and transketolase-like-2 genes) between the transfected human colon cancer cell line cells and the controls (P > 0.05). Our data demonstrated that the transketolase-like-1 gene plays an important role in total transketolase activity and in the cell proliferation of human colon cancer. Transketolase-like-1 may serve as a target for novel anticancer therapies.