摘要
目的构建STAT3基因的SiRNA表达载体,并观察其对小细胞肺癌细胞增殖能力的影响。方法根据STAT3序列设计SiRNA引物,构建SiRNA表达载体(STAT3-SiRNA-1,STAT3-SiRNA-2,STAT3-SiRNA-N)。应用Lipofect2000将STAT3-SiRNA表达载体转染NCI-H446细胞,流式细胞仪检测STAT3基因表达,噻唑兰法检测转染STAT3基因对小细胞肺癌细胞增殖能力的影响。结果STAT3载体构建成功。转染STAT3-SiRNA-1、STAT3-SiRNA-2后,STAT3基因表达明显下降;转染STAT3-SiRNA-1、STAT3-SiRNA-2的细胞增殖能力明显低于转染STAT3-SiRNA-N和未转染的细胞(P<0.05)。结论成功构建了STAT3-SiRNA表达载体,该载体能有效抑制小细胞肺癌细胞STAT3基因的表达,与小细胞肺癌增殖活性有关。
Objective Construct SiRNA expression vector and observe the influence of SiRNA expression vector on small cell lung carcinoma cells proliferation. Methods We designed SiRNA primer by STAT3 sequence and constructed SiRNA expression vector （STAT3-SiRNA-1, STAT3-SiRNA-2, STAT3-SiRNA-N）. STAT3-SiRNA expression vector were transfected into NCI-Hd46 cells with lipofect2000. The STAT3 expression was detected by flow cytometry. The influence of STAT3 gene on the proliferation of small cell lung cancer cells was assayed by MTT methods. Results SiRNA vector of STAT3 proved to be successfully constructed. After transfected with STAT3-SiRNA-1 or STAT3-SiRNA-2, the expression of STAT3 gene was obviously decreased. The proliferation of the cells which transfected with STAT3-SiRNA-1, STAT3-SiR- NA-2 were much lower than that of cells which were transfected with STAT3-SiRNA-N and no transfection cells. Conclusions STAT3-SiRNA expression vector is successfully constructed, which can effectively inhibit the expression of STAT3 gene in NCI-H446 cells. STAT3 gene may relate with the proliferation of small cell lung cancer.
出处
《山东医药》
CAS
北大核心
2008年第34期24-26,共3页
Shandong Medical Journal
基金
辽宁省教育厅资助项目(2004D162)。
