HSU17714基因反义重组质粒的构建及其对人大肠癌细胞生长的影响

Construction of Recombinant HSU17714 Gene Antisense Expressive Plasmid and Its Influence on Colon Cancer Cells Growth

我所应用减式杂交方法筛选人大肠癌负相关基因,得到一与已知基因同源性仅50％的cDNA片段,已作为新发现的基因被NCBI收录入国际基因库,我们将此cDNA片段反向克隆至非整合型哺乳动物表达载体pREP9的多克隆位点中,构建了HSU17714基因的反义RNA表达载体,并籍脂质体将之导入人结肠癌SW1116细胞中,双层软琼脂集落培养试验、流式细胞技术分析及MTT比色法等结果分别提示转染该重组体的肠癌细胞的非锚着依赖性生长能力、增殖周期活性和细胞生长速度均受到不同程度的抑制,说明外源重组体pREP9+HSU17714(AS)对肠癌SW1116细胞的生长具有抑制作用。

HSU17714 gene is a novel human colorectal carcinoma related gene isolated by subtractive hybridization method. The recombinant antisense expressive plasmid of HSU17714 gene was constructed by inserting the partial sequence (2612 bp) of this gene into the multiple cloning site of the vector pREP9 inversely. Then, it was transferred into human colon adenocarcinoma cells (SW1116) through liposome mediation. Obtained from the two-layer soft agarose culture test, flowcytometry test and cell growth rate detection, the data indicated that the antisense HSU17714 could inhibit the growth of SW1116 cells at different degrees. So, a conclusion could be drawn that the antisense HSU17714 gene expressive vector plays a role in the inhibition of SW1116 cell growth.