中文摘要：

目的：构建针对M2型丙酮酸激酶（PKM2）基因3个可干扰序列设计小发夹RNA（small hairpin RNA,shRNA）载体,为下一步探索Ad-PKM2介导PKM2 RNAi对乳腺癌细胞作用及机制建立基础。方法：根据RNAi设计软件设计并合成3对shRNA模板序列,依次将它们克隆至pGenesil载体中构建重组质粒,通过MTT检测筛选出对乳腺癌细胞株MCF-7增殖效应影响最强的RNAi序列。结果：酶切鉴定和测序结果证实RNAi载体构建成功,均能高效感染MCF-7并抑制MCF-7细胞的增殖,pGenesil-PKM2-（1＋2＋3）的抑制能力最强。结论：针对PKM2多靶点RNAi载体成功构建,为下一步研究PKM2被沉默后细胞生物学行为的变化打下基础。

英文摘要：

Objective： To construct a small hairpin RNA（ shRNA） expression vector,mediated by recombinant adenovirus and targeting PKM2,and which will help for exploring the biological behavior change of breast cancer cells by means of knockingdown PKM2. Methods： Three pairs of templates of PKM2 shRNA were synthesized and cloned into the pGenesil system successfully. Then its effect on the proliferation of MCF-7 was detected by MTT. Results： pGenesil-PKM2 was constructed successfully,and also it repressed proliferation of MCF-7 significantly after being transfected into the cells. Conclusion： The construction of pGenesil-PKM2 paved the way for the further research.