山东医药2011，Vol.51Issue(8)：13-16,4.
Smoothened shRNA载体的构建及其对乳腺癌细胞增殖的影响
Construction of Smoothened shRNA expression vector and effects on the proliferation of breast cancer cells
摘要
Abstract
Objective To construct and screen the RNAi expression vector for Smoothened (SMO) gene and explore the effects of Hedgehog signaling on the proliferation of breast cancer cells. Methods Breast cancer MCF-7 cells were transfected for 48 hours by the mediation of LipofectamineTM 2000. The transcription level of SMO mRNA and expression of SMO protein in transfected cells detected by realtime RT-PCR and Western blot methods. CCK-8 kit and real time RT-PCR were employed to study the effects of SMO mRNA transfection on the proliferation of breast cancer cells and cyclinD1 mRNA. Results It was verified by partial nucleotide sequencing that the constructed eukaryotic vectors expressing shRNA of SMO were correct. Contrast to MOCK group, the SMO mRNA expression in SMO shRNA-1 group, SMO shRNA-3 group,SMO shRNA-4 group declined (P= 0. 015,0. 002, 0. 000), moreover the SMO mRNA expression in SMO shRNA-4 group obviously lower than other 3 groups (P =0. 007, 0. 000, 0. 046). The inhibitor rate of SMO shRNA-4 was 87％ , meanwhile the expression of SMO protein decreased. MCF-7 cell proliferation and cyclinD1 mRNA significantly inhibited under the condition of transfected by SMO shRNA-4. Conclusions The RNAi expression vector for SMO gene is successfully constructed and screened. SMO shRNA visibly inhibite the proliferation of breast cancer cells. Hedgehog signaling can induce cell proliferation by actived cyclinD1.关键词
Smoothened/shRNA/载体构建/增殖/cyclinD1分类
医药卫生引用本文复制引用
陶雅军,毛俊,张晴晴,李连宏..Smoothened shRNA载体的构建及其对乳腺癌细胞增殖的影响[J].山东医药,2011,51(8):13-16,4.基金项目
国家自然科学基金资助项目(30870968) （30870968）
教育部高等学校博士学科点专项科研基金(200801610001). （200801610001）
