摘要
目的探讨聚乙烯亚胺(PEI)介导的人蛋白激酶Cα(PKCα)基因反义寡核苷酸(ASODN)对人肺癌A549细胞生长增殖及PKCα基因表达的影响。方法以PEI介导PKCα ASODN,随机寡核苷酸(RODN)分别转染A549细胞,采用WST法和克隆形成抑制实验分析细胞生长增殖抑制效果;RT-PCR检测PKCα mRNA的表达水平;Western blotting检测PKCα蛋白的表达水平。结果PEI介导的PKCα ASODN转染A549细胞后,细胞生长增殖和克隆形成均受到抑制(P<O.05),并呈浓度依赖关系,其抑制率明显高于PEI或PEI介导的RODN组;RT-PCR和Western blotting结果均表明:PEI介导的PKCα ASODN转染A549细胞能明显抑制PKCα的表达,与空白对照组、PEI或PEI介导的RODN转染组相比,有统计学差异(P<0.05)。结论PEI介导的PKCα反义核酸能下调PKCα基因的表达,显著抑制A549细胞生长和增殖。
Objective To investigate the effects of antisense oligonucleotides （ASODN） targeting protein kinase C α （PKCα） on the proliferation ofA549 cells. Methods PKCα ASODN and random oligonucleotides （RODN） were transfected into A549 cells mediated by polyethyleneimine, and the proliferation and clone formation ofA549 cells were detected by CCK-8 and clone formation assay, respectively. The expression of PKCα in the transfected cells was analyzed by RT-PCR and Western blotting. Results Compared with those in the control group, PEI group and PEI-RODN group, the proliferation and clone formation of A549 cells treated with ASODN targeting PKCα were significantly inhibited （P〈0.05）. The expressions of PKCα mRNA and protein in PKCα ASODN-transfected A549 cells were significantly lower than those in the other 3 groups （P〈0.05）. Conclusion The PKCα ASODN mediated by PEI down-regutates the expression of PKCα gene and suppress the proliferation and clone formation of A549 cells.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2009年第6期1189-1191,1203,共4页
Journal of Southern Medical University
基金
暨南大学临床学院科研培育基金(暨医临2005-05)
关键词
蛋白激酶CΑ
反义核酸
A549
聚乙烯亚胺
protein kinase C α
antisense oligodeoxynucleotide
A549 cells
polyethyleneimine
