microRNA-592对人结直肠癌细胞增殖迁移的影响

Effect of MicroRNA- 592 on Proliferation and Migration of Human Colorectal Neoplasm Cells

目的探讨microRNA-592(miR-592)在结直肠癌细胞系中的表达并观察对结直肠癌细胞系增殖、迁移的影响。方法 Real-time PCR检测miR-592在四种结直肠癌细胞系与正常结直肠细胞系的表达差异;构建miR-592的抑制表达载体并包装慢病毒,转染Lovo细胞,筛选出稳定表达的细胞株并Real-time PCR检测Ki67、Cyclin D1、P27的mRNA的表达;MTT、平板克隆和划痕实验分别检测miR-592对Lovo细胞增殖和迁移的影响。结果 miR-592在四种结直肠癌细胞系(HCT116、Lovo、LS174T、SW480)中的表达与正常结直肠细胞系CCD-18Co相比均明显增高(P均<0.01)。稳定转染后,Ki67、Cyclin D1的mRNA在Lovo细胞中的表达水平明显降低,P27mRNA的水平明显升高;Lovo细胞生长减慢,克隆形成减少,细胞迁移能力降低(P均<0.05或<0.01)。结论 miR-592在结直肠癌细胞中呈高表达,抑制miR-592表达可降低Lovo细胞的增殖和迁移能力。

Objective To explore the expression of microRNA- 592（ miR- 592） in colorectal neoplasm cell lines and the effect of miR- 592 on the proliferation and migration of colorectal neoplasm cell lines. Methods The expression of miR- 592 in different colorectal neoplasm cell lines and normal colorectal cell line was examined by quantitative real- time PCR. The lentiviral inhibited expression vector of miR- 592 and the lentiviral negative control vector were constructed and transfected into Lovo cells,Lovo cell lines with stable expression of miR- 592 or control vectors were obtained in medium supplemented by puromycin,designated Lovo- sh592 and Lovo- NC,respectively. The mRNA expression of Ki67,Cyclin D1 and P27 were examined by quantitative real- time PCR in Lovo- sh592 and Lovo- NC cells. Cell proliferation was measured in vitro by MTT assay and tablet cloning experiments,cell migration was measured in vitro by wound healing assay.Results Compared with normal colorectal cell line CCD- 18 Co,the expression of miR- 592 in four colorectal neoplasm cell lines（ HCT116,Lovo,LS174 T,SW480） significantly increased. In HCT116 cells the expression of miR- 592 was（ 2. 98 ± 0. 37）（ P〈0. 05）,in Lovo cells the expression of miR- 592 was（ 10. 65 ±0. 27）（ P〈0. 05） and in LS174 T cells the expression of miR- 592 was（ 126. 47 ± 0. 46）（ P〈0. 01）,in SW480 cells the expression of miR- 592 was（ 7. 61 ± 0. 18）（ P〈0. 05）. After stable transfection,compared with Lovo- sh592 cells,the mRNA expression of Ki67 and Cyclin D1 significantly decreased,and the expression of P27 significantly increased in Lovo- NC cells. After the expression of miR- 592 was inhibited,the Lovo cell growth shown to be significantly slow. The colony formation and cell migration were reduced compared with control group. Conclusion The expression of miR- 592 is upregulated in human colorectal neoplasm. Low expression of miR- 592 can significantly inhibit Lovo cell proliferation and migration. miR- 592 may be used as ＂oncogene＂participation in colorectal neoplasm incidence and progression.