摘要
目的:探讨Dkkl cDNA抗结肠癌的机制.方法:利用脂质体介导pcDNA3.1(+)Dkk1进行瞬时转染结肠癌细胞CT26和表达Dkk1的肝癌细胞HEPA1-6,转染后的细胞为实验组,转染空载体pcDNA3.1(+)的细胞和未转染的细胞分别为空载体组和未转染组,用MTT法检测CT26增殖、免疫印迹法检测细胞Dkk1,p53,Bcl-2和Bax的表达量.结果:MTT实验显示在570 nm的吸光度均值CT26实验组明显低于其空载体组和未转染组(0.779±0.025 vs 0.968±0.016,0.973±0.016),P<0.05);与CT26空载体组和未转染组相比,实验组p53,Bcl-2的表达量降低、而Dkk1、Bax表达增高.结论:外源Dkk1可反馈抑制突变型p53的生成,下调Bcl-2,增加Bax因子表达而抑制结肠癌的增殖.
AIM： To clone the cDNA of Dickkopf-1 （Dkk1） and to investigate its anti-colon carcinoma activity.
METHODS： Dkk1 cDNA was retrived from Mouse Embryo 9 dpc, DH10B cDNA library, and eukaryotic expression vector pcDNA3.1（＋） Dkk1 was constructed. Transfected CT26 colon carcinoma cells served as the experimental group, transfecfion empty vector pcDNA3.1（＋） cells served as the empty vector group, and nontransfected cells as the non-transfected group. Proliferation of the CT26 colon carcinoma cells was measured by MTT assay. Expression of Dkk1, p53, Bcl-2 and Bax was detected by Western blotting.
RESULTS： Absorbance at 570 nm in the experimental group was lower than that in the empty vector and non-transfected groups （0.779±0.025 vs 0.968±0.016 and 0.973±0.016, P 〈 0.05）. Compared with the empty vector and nontransfected groups, the level of p53 and Bcl-2 in CT26 cells was lower, and the level of Dkk1 and Bax was higher.
CONCLUSION： Inhibition of CT26 cell proliferation by Dkk1 expression may be mediated by Dkk1 blocking mutant p53, decreasing Bcl-2 and enhancing Bax expression.
出处
《世界华人消化杂志》
CAS
北大核心
2008年第1期20-24,共5页
World Chinese Journal of Digestology
作者简介
李甲初，重庆医科大学生物化学与分子生物学硕士生，主要从事肿瘤分子生物学方面的研究．
通讯作者：曾昭淳，400016，重庆市渝中区医学院路1号，重庆医科大学生物化学与分子生物学教研室，重庆市生物化学与分子药理重点实验室．zengzc88@yahoo．com．cn电话：023-68485398
