Vector-based miR-15a/16-1 plasmid inhibits colon cancer growth in vivo
Vector-based miR-15a/16-1 plasmid inhibits colon cancer growth in vivo
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DOI：
10.1042/cbi20110404
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发表时间：
2012-08-01
影响因子：
3.9
通讯作者：
Deng, Hong-Xin
中科院分区：
文献类型：
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作者：
Dai, Lixia;Wang, Wei;Deng, Hong-Xin
miR-15（microRNA 15）和miR-16在多种肿瘤细胞系和多种肿瘤组织中频繁缺失或下调，表明miR-15 a/16-1在肿瘤进展中起重要作用，并可能成为癌症治疗的方法。我们已经开发了一种基于载体的质粒来探索miR-15 a/16-1在体内结肠癌中的抗肿瘤功效。据推测，miR-15 a和miR-16-1靶向细胞周期蛋白B1（CCNB 1），其与几种致瘤特征如存活和增殖相关。miR-15 a和miR-16-1在结肠癌细胞中的表达水平与CCNB 1的表达呈负相关，同源性分析发现miR-15 a/16-1与CCNB 1的mRNA序列具有一致性。构建miR-15 a/16-1表达载体，体外转染HCT 116和SW 620结肠癌细胞。使用流式细胞术分析、集落形成分析和管形成分析来分析对细胞活力和血管生成产生的影响。Western blotting检测CCNB 1表达下调。包裹在阳离子脂质体中的miR-15 a/16-1质粒的全身递送导致皮下肿瘤生长和肿瘤组织中的血管生成的显著抑制，而使用携带非特异性质粒的脂质体没有观察到效果。综上所述，miR-15 a/16-1已被应用于结肠癌的体内治疗，并导致结肠肿瘤异种移植物的有效生长停滞和血管生成减少。这些发现表明，基于载体的miR-15 a/16-1表达质粒的系统递送可以成为结肠癌治疗的一种方法。
miR-15 (microRNA 15) and miR-16 are frequently deleted or down-regulated in many cancer cell lines and various tumour tissues, suggesting that miR-15a/16-1 plays important roles in tumour progression and might be a method for cancer treatment. We have developed a vector-based plasmid to explore the anti-tumour efficacy of miR-15a/16-1 in colon cancer in vivo. It is proposed that miR-15a and miR-16-1 target cyclin B1 (CCNB1), which associates with several tumorigenic features such as survival and proliferation. The levels of miR-15a and miR-16-1 in colon cancer cells were inversely correlated with CCNB1 expression, and there was consensus between miR-15a/16-1 and CCNB1 mRNA sequences by analysing homology. Vector-based miR-15a/16-1 expression plasmid was constructed and transfected into HCT 116 and SW620 colon cancer cells in vitro. The effects produced on cell viability and angiogenesis were analysed using flow cytometric analysis, colony formation analysis and tube formation analysis. CCNB1 expression down-regulation was checked by Western blotting. Systemic delivery of miR-15a/16-1 plasmids encapsulated in cationic liposome led to a significant inhibition of subcutaneous tumour growth and angiogenesis in tumour tissues, whereas no effects were observed with liposome carrying the non-specific plasmid. In summary, miR-15a/16-1 has been applied in colon cancer treatment in vivo, and resulted in effective colon tumour xenografts growth arrest and angiogenesis decrease. These findings suggest that systemic delivery of vector-based miR-15a/16-1 expression plasmid can be an approach to colon cancer therapy.