论文部分内容阅读
目的:运用RNA干扰技术,观察siRNA表达载体在乳腺癌细胞MDA-MB-231中对CaSR基因表达的影响。方法:构建靶向CaSR基因的RNA干扰表达载体,用脂质体转染乳腺癌细胞株MDA-MB-231,运用Real-Time荧光定量PCR,Westernblot技术分别从mRNA以及蛋白表达水平检测CaSR基因表达的变化。结果:所构建的质粒载体成功的在MDA-MB-231细胞中抑制了CaSR mRNA及其蛋白的表达。与对照组相比,psiRNA-CaSR载体对CaSR mRNA的抑制率达到65%,对CaSR蛋白抑制率约为70%。结论:实验证明所设计的shRNA片段可以有效地抑制CaSR基因的表达,为下一步研究工作奠定了基础。
Objective: To observe the effect of siRNA expression vector on CaSR gene expression in breast cancer cell MDA-MB-231 using RNA interference technology. METHODS: RNA interference expression vector targeting CaSR gene was constructed and transfected into breast cancer cell line MDA-MB-231 by lipofectamine. Real-time fluorescence quantitative PCR and Western blot were used to detect CaSR gene expression from mRNA and protein expression levels, respectively. The change. Results: The constructed plasmid vector successfully inhibited CaSR mRNA and protein expression in MDA-MB-231 cells. Compared with the control group, the inhibition rate of CaSR mRNA in the psiRNA-CaSR vector reached 65%, and the inhibition rate to CaSR protein was approximately 70%. Conclusion: Experiments show that the designed shRNA fragment can effectively inhibit the expression of CaSR gene, which lays a foundation for the next research work.