Huang Hu, Hu Yi-de, Li Na, Zhu Yong
Third Department of Oncology, XinQiao Hospital, Third Military Medical University, ChongQing, PR China.
Oligonucleotides. 2009 Jun;19(2):151-62. doi: 10.1089/oli.2008.0174.
To observe whether cyclin D1 siRNA-mediated inhibition of cyclin D1 represents a promising antigrowth and antimetastatic strategy for cancer gene therapy, particularly for non-small cell lung cancers. To stably transfect the A549 cell line with a cyclin D1-targeted siRNA to downregulate cyclin D1 expression and observe the effects on protein expression, and tumor growth in vitro and in vivo. Expression of cyclin D1-targeted siRNA resulted in a decrease in cyclin D1, MMP-2, RhoA, and Rac1 protein levels, as detected by Western blot and immunofluorescence studies. Transfected cells also exhibited a marked decrease in the rate of cell growth, and decreased invasive capacity, compared to cells transduced with a scrambled siRNA plasmid and untransduced A549 cells. siRNA-mediated inhibition of cyclin D1 expression represents a promising antigrowth and antimetastatic strategy for cancer gene therapy, particularly for non-small cell lung cancers. It is the reason for inhibiting tumor growth so that cyclin D1 siRNA can inhibit the cell cycle progression. In addition, the mechanism of inhibiting tumor metastasis was related to the decrease in the expression of MMP-2, RhoA, and Rac1 after cyclin D1 was decreased by cyclin D1 siRNA.
观察细胞周期蛋白D1小干扰RNA(siRNA)介导的细胞周期蛋白D1抑制作用是否代表一种有前景的癌症基因治疗抗生长和抗转移策略,尤其是对于非小细胞肺癌。用靶向细胞周期蛋白D1的siRNA稳定转染A549细胞系,以下调细胞周期蛋白D1表达,并观察其对蛋白质表达以及体外和体内肿瘤生长的影响。通过蛋白质免疫印迹和免疫荧光研究检测发现,靶向细胞周期蛋白D1的siRNA表达导致细胞周期蛋白D1、基质金属蛋白酶-2(MMP-2)、RhoA和Rac1蛋白水平降低。与用乱序siRNA质粒转导的细胞和未转导的A549细胞相比,转染细胞的细胞生长速率也显著降低,侵袭能力下降。siRNA介导的细胞周期蛋白D1表达抑制代表一种有前景的癌症基因治疗抗生长和抗转移策略,尤其是对于非小细胞肺癌。这就是抑制肿瘤生长的原因,即细胞周期蛋白D1 siRNA可抑制细胞周期进程。此外,抑制肿瘤转移的机制与细胞周期蛋白D1 siRNA使细胞周期蛋白D1降低后MMP-2、RhoA和Rac1表达下降有关。