沉默KDR基因乳腺癌MCF-7细胞的生物学特征改变

KDR gene silencing effect on biological properties of breast cancer MCF-7 cells

背景:随着基因工程以及肿瘤生物分子学等新兴学科的发展壮大,基因治疗肿瘤成为一种新的治疗模式。目的:探讨KDR基因沉默对乳腺癌MCF-7细胞增殖能力和侵袭能力的影响。方法:设计针对KDR基因的小分子干扰RNA(siR NA)序列,转染人乳腺癌MCF-7细胞,应用RT-PCR及Western blot法检测沉默KDR基因后MCF-7细胞KDR mR NA及蛋白的表达;应用流式细胞仪、CCK-8增殖实验、小室侵袭实验检测沉默KDR基因后乳腺癌MCF-7细胞的细胞周期、增殖能力、侵袭能力的变化。结果与结论:KDR基因沉默48 h后,乳腺癌MCF-7细胞KDR mR NA及蛋白表达明显降低;乳腺癌MCF-7细胞停滞在G0/G1周期,S期的细胞数目减低,细胞增殖得到显著抑制,穿过滤膜的细胞数量明显减少。上述结果表明沉默KDR基因后乳腺癌MCF-7细胞的增殖、侵袭能力得到明显抑制,说明KDR基因沉默可能成为新的有效治疗乳腺癌的靶点。

BACKGROUND：With the development of genetic engineering and tumor molecular biology, gene therapy for tumors has become a new treatment modality. OBJECTIVE：To explore the effect of the KDR gene silencing on the proliferation and invasion capacity of breast cancer MCF-7 cel s. METHODS：Interfering RNA （siRNA） sequences for smal molecule KDR gene was designed and transferred into human breast cancer MCF-7 cel s. Then, RT-PCR and western blot assay were used to detect the KDR mRNA and protein expression. Flow cytometry, cel counting kit-8 test and Transwel test were employed to detect the cel cycle, proliferative capacity and invasion capacity of breast cancer MCF-7 cel s after the KDR gene silencing. RESULTS AND CONCLUSION：After 48 hours of KDR silencing, the mRNA and protein expressions of KDR in MCF-7 cel s were decreased obviously;MCF-7 cel s arrested at G0/G1 stage and the number of cel s at S stage was reduced. Cel proliferation was inhibited significantly. The amount of cel s passing through the filtering membrane became less. After KDR gene silencing, the proliferation and invasion of breast cancer MCF-7 cancer stem cel s were inhibited remarkably, indicating that KDR gene silencing may be a new target for the effective treatment of breast cancer.