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目的 :探讨靶向沉默Xklp2靶蛋白(targeting protein for Xenopus kinesinlike protein 2,TPX 2)基因对结肠癌SW480细胞增殖、侵袭的影响及其可能的作用机制。方法:将靶向沉默TPX2基因的重组载体p Magic4.1-sh RNA-TPX2转入结肠癌SW480细胞后,应用MTT法检测SW480细胞的增殖能力,Transwell小室法检测细胞的侵袭能力,RT-PCR和蛋白质印迹法检测SW480细胞中TPX2、血管内皮生长因子(vascular endothelial growth factor,VEGF)和基质金属蛋白酶9(matrix metalloproteinase-9,MMP-9)mRNA及蛋白的表达。结果 :重组载体p Magic4.1-shRNA-TPX2转染后,SW480细胞的增殖和侵袭能力均显著低于阴性对照组(阴性对照载体p Magic4.1-sh RNA-NC转染至SW480细胞)和空白对照组(未进行任何转染的SW480细胞)(P值均<0.01);p Magic4.1-shRNA-TPX2转染组SW480细胞中TPX2、VEGF、MMP-9 m RNA及蛋白的表达水平均显著低于阴性对照组和空白对照组(P值均<0.01)。结论 :靶向沉默TPX2基因能够显著抑制结肠癌SW480细胞的增殖和侵袭,这一作用可能与VEGF和MMP-9基因的表达下调有关。
OBJECTIVE: To investigate the effect of target protein targeting Xenopus kinesin-like protein 2 (TPX2) on proliferation and invasion of colon cancer SW480 cells and its possible mechanism. Methods: The recombinant vector p Magic4.1-sh RNA-TPX2 targeting silencing TPX2 gene was transfected into SW480 cells. MTT assay was used to detect the proliferation of SW480 cells. The invasion ability of SW480 cells was detected by Transwell chamber assay. RT-PCR Western blotting was used to detect the mRNA and protein expressions of TPX2, VEGF and MMP-9 in SW480 cells. Results: The proliferation and invasion ability of SW480 cells were significantly lower than that of the negative control group (negative control vector p Magic4.1-sh RNA-NC transfected SW480 cells) and The expression of TPX2, VEGF, MMP-9 mRNA and protein in SW480 cells transfected with p Magic4.1-shRNA-TPX2 were significantly higher than those in control group (all SW480 cells without any transfection) Significantly lower than the negative control group and the blank control group (P value <0.01). CONCLUSION: Targeting silencing of TPX2 gene can significantly inhibit the proliferation and invasion of colon cancer SW480 cells, which may be related to the down-regulation of VEGF and MMP-9 gene expression.