摘要
目的观察靶向S100A4的shRNA对乳腺癌MCF-7细胞增殖的影响。方法构建S100A4基因特异性shRNA表达载体,检测转染后MCF-7细胞中S100A4的表达水平以及增殖水平和凋亡率的变化。经过脂质体介导将S100A4shRNA表达载体转染入MCF-7细胞,G418筛选及克隆化培养,裸鼠体内成瘤试验检测各组细胞增殖情况。结果经测序鉴定证实成功构建S100A4-shRNA表达载体。所构建的S100A4-shRNA表达载体能够有效抑制MCF-7细胞中S100A4的表达,并且可以显著降低MCF-7细胞增殖水平以及诱导细胞进入晚期凋亡;稳定转染S100A4-shRNA表达载体的MCF-7细胞形态无显著变化,而实验组裸鼠肿瘤体积和重量明显小于空白对照组和阴性对照组。结论S100A4-shRNA表达载体能有效地抑制乳腺癌MCF-7细胞中S100A4的表达,从而降低细胞增殖水平。
Objective To study the effect of short hairpin RNA （shRNA）- targeted St00A4 on proliferation of breast cancer MCF-7 cells. Methods S 100A4-shRNA expression vector, constructed and confirmed by sequencing, was transfected into MCF-7 cells using lipofectamine ^TM2000. Expression, proliferation and apoptosis of S 100A4 in MCF-7 cells were detected by quantitative RT- PCR and Western blot 48h after transfection. S100A4-shRNA expression vector was transfected into breast cancer MCF-7 cells and G418 was selected and cloned. Proliferation of breast cancer MCF-7 cells in nude mice was detected by in vivo tumor formation test. Results S100A4-shRNA expression vector was constructed and transfected into MCF-7 cells as confirmed by sequencing. The constructed vector could effectively inhibit the expression of S100A4 in MCF-7 cells, decrease the proliferation of MCF-7 cells, and induce the cells into late apoptosis. No significant change in MCF-7 cells was observed after stable transfection of S100A4 mRNA expression vector. However, the volume and weight of tumor were significantly decreased compared with the control group. Conclusion S 100A4-shRNA expression vector can significantly suppress S 100A4 expression and inhibit the proliferation of MCF-7 cells.
出处
《军医进修学院学报》
CAS
2010年第1期63-66,共4页
Academic Journal of Pla Postgraduate Medical School
基金
广东省科技计划资助 (2008B030301345)~~
