Leng Aimin, Yang Jing, Liu Ting, Cui Jianfang, Li Xiu-Hua, Zhu Yanan, Xiong Ting, Chen Yuxiang
Department of Gastroenterology, Xiangya Hospital, Changsha, 410008, China.
Tumour Biol. 2011 Dec;32(6):1103-11. doi: 10.1007/s13277-011-0210-5. Epub 2011 Jul 15.
The strategies for tumor-specific expression of suicide genes and target tumor angiogenesis have been tested in tumors. However, the anti-tumor efficacy of the combination of these two strategies, particularly, delivering suicide gene and anti-angiogenesis agent by nanoparticles, has not yet been evaluated in colon carcinoma. We constructed a cassette to silence VEGF-A expression and express a fused yCDglyTK gene driven by tumor-specific promoter (shVEGF-CDTK). The DNA carrying shVEGF-CDTK was delivered into colon carcinoma cells by calcium phosphate nanoparticles (CPNPs). Cell proliferation was measured by MTT assay, and apoptosis was detected by flow cytometry. The anti-tumor effect of the combined cassette was tested in xenograft animal model. With 5-fluorocytosine (5-FC), CPNP-delivered shVEGF-CDTK DNA (CPNP-shVEGF-CDTK) showed high expression of fused yCDglyTK gene and effectively silenced VEGF-A expression in vitro and in vivo, which significantly inhibited colon carcinoma cell proliferation and induced apoptosis in vitro. With 5-FC, the systemic delivery of CPNP-shVEGF-CDTK significantly inhibited tumor growth in the colon carcinoma xenograft animal model. The combined cassette is obviously effective in inhibiting tumor cell proliferation and inducing apoptosis in vitro and tumor growth in vivo than the CPNP-shVEGF or CPNP-CDTK alone. The combination of VEGF-A-silencing and tumor-specific expression of suicide gene is an effective strategy for colon carcinoma treatment.
自杀基因的肿瘤特异性表达策略以及靶向肿瘤血管生成的策略已在肿瘤中进行了测试。然而,这两种策略联合使用的抗肿瘤效果,特别是通过纳米颗粒递送自杀基因和抗血管生成剂的效果,尚未在结肠癌中得到评估。我们构建了一个盒式结构,以沉默VEGF-A的表达并表达由肿瘤特异性启动子驱动的融合yCDglyTK基因(shVEGF-CDTK)。携带shVEGF-CDTK的DNA通过磷酸钙纳米颗粒(CPNP)递送至结肠癌细胞。通过MTT法测量细胞增殖,并通过流式细胞术检测细胞凋亡。在异种移植动物模型中测试了联合盒式结构的抗肿瘤作用。与5-氟胞嘧啶(5-FC)联合使用时,CPNP递送的shVEGF-CDTK DNA(CPNP-shVEGF-CDTK)在体外和体内均显示出融合yCDglyTK基因的高表达,并有效沉默了VEGF-A的表达,这在体外显著抑制了结肠癌细胞的增殖并诱导了细胞凋亡。与5-FC联合使用时,CPNP-shVEGF-CDTK的全身递送在结肠癌异种移植动物模型中显著抑制了肿瘤生长。联合盒式结构在体外抑制肿瘤细胞增殖和诱导细胞凋亡以及在体内抑制肿瘤生长方面明显比单独的CPNP-shVEGF或CPNP-CDTK更有效。沉默VEGF-A与自杀基因的肿瘤特异性表达相结合是结肠癌治疗的有效策略。