慢病毒介导含菱形结构蛋白1基因下调抑制结肠癌细胞增殖的研究

Lentivirus-mediated rhomboid domain containing 1 knockdown inhibits colorectal cancer cell growth

目的 探讨含菱形结构蛋白1（RHBDD1）基因对结肠癌细胞增殖影响及机制.方法 使用慢病毒介导RNA干扰技术下调RKO细胞RHBDD1基因水平;噻唑蓝（MTT）法、克隆形成实验检测RKO细胞增殖变化;流式细胞仪检测细胞周期变化.结果 实时定量聚合酶链反应（Real-time PCR）检测Lv-shRHBDD1组荧光强度（0.49±0.35）明显降低,证实RNA干扰下调了RHBDD1基因水平（P＜0.01）;MTT法显示RHBDD1基因下调后RKO细胞增殖率显著低于未下调细胞（P＜0.01）;克隆形成试验显示Lv-shRHBDD1组RKO细胞克隆数（35.0±2.9）明显减少（P＜0.01）;流式细胞仪分析显示Lv-shRHBDD1组G0/G1期[（50.20±1.22）％]细胞比例增多,S期[（39.30±0.87）％]和G/M期[（8.10±0.72）％]细胞比例下降,表明慢病毒介导下调RHBDD1基因的RKO细胞周期阻滞于G0/G1期（P＜0.01）.结论 RHBDD1在结肠癌细胞增殖过程中发挥重要作用.

Objective To investigate the role of rhomboid domain containing 1 （RHBDD1） in colorectal carcinoma （CRC） tumorigenesis.Methods Lentivirus-mediated RNA interference （RNAi） was employed to knock down RHBDD1 in RKO cells.RKO cell proliferation alteration was examined by methyl thiazol tetrazolium （MTT） and colony formation capacity with RKO monolayer cell culture was assessed;Cell cycle analysis was performed by flow cytometry.Results Real-time quantitative polymerase chain reaction （Real-time PCR） showed arbltrary unit of Lv-shRHBDD1 group decreased （0.49 ± 0.35）,which confirmed knockdown efficiency （P ＜0.01）.Functional analysis indicated that depletion of RHBDD1 could significantly down-regulate CRC cell proliferation （P ＜ 0.01）.Colony formation capacity of Lv-shRHBDD1 group （35.0 ± 2.9） was significantly reduced （P ＜ 0.01）.Flow cytometry showed that RKO cell proportion of Lv-shRHBDD1 group was increased in G0/G1 phase （35.0 ± 2.9）,and decreased in S phase [（39.30 ± 0.87） ％] and G2/M phase [（8.10 ± 0.72） ％] （P ＜ 0.01）.Conclusion It could be concluded from this study that RHBDD1 might contribute to CRC tumorigenesis and serve as a potential therapeutic target in human colon cancer.