KOZAK序列对乳腺癌MDA-MB-231细胞CCNL1基因表达的影响

Enhancement of cyclin L1 expression by a key sequence-KOZAK in MDA-MB-231 cells

目的:探讨KOZAK序列对细胞周期调节蛋白1(CCNL1)基因在人乳腺癌细胞MDA-MB-231中表达的影响。方法:构建pcDNA3.1-CCNL1和pcDNA3.1-CCNL1-K(含有KOZAK序列)的真核重组表达质粒,采用Fugene HD转染试剂将重组表达质粒转染入MDA-MB-231细胞,并用荧光定量PCR和Western blot方法检测pcDNA3.1-CCNL1和pcDNA3.1-CCNL1-K重组质粒在MDA-MB-231细胞中表达的差异。结果:在MDA-MB-231细胞中质粒pcDNA3.1-CCNL1-K比空白对照细胞mRNA和蛋白质表达量要高;而质粒pcDNA3.1-CCNL1比空白对照细胞mRNA和蛋白质表达量也高。结论:在MDA-MB-231细胞中,质粒pcDNA3.1-CCNL1-K较pcDNA3.1-CCNL1的mRNA和蛋白表达水平均存在差异,KOZAK序列能够提高CCNL1基因在MDA-MB-231细胞中的表达。

Objective： To explore whether the expression of CCNL1 gene and the KOZAK sequence can enhance the expression of CCNL1 gene in breast cancer cells（MDA-MB-231）.Methods：The eukaryotic expression plasmids（pcDNA3.1-CCNL1 and pcDNA3.1-CCNL1-K） were constructed.They were transfected into the MDA-MB-231 cells by Fugene HD transfection reagents.Detect the expression of pcDNA3.1-CCNL1 and pcDNA3.1-CCNL1-K in the MDA-MB-231 cells by real time PCR and Western blot.Results：The KOZAK sequence could enhance the expression of mRNA and protein in the MDA-MB-231 cells.Conclusion：KOZAK sequence play a key role in CCNL1 protein expression.