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目的结肠癌的侵袭和转移是结肠癌患者死亡的主要原因。文中拟探讨叉头框蛋白M1(Forkhead box M1,FoxM1)对人结肠癌细胞恶性表型的影响。方法应用实时荧光定量PCR(real time-PCR,RT-PCR)和Western blot方法筛选出高表达细胞株HT-29及低表达细胞株HCT-116,应用RNA干扰技术有效下调FoxM1在HT-29细胞中的表达,同时采用过表达质粒调高FoxM1在HCT-116中的表达,2种细胞株根据不转染、转染空载质粒及转染干扰或过表达FoxM1质粒各分为3组:空白对照组、实验对照组、实验组。分别采用划痕实验和Transwell小室法检测上述转染细胞的细胞增殖、迁移与侵袭能力的变化。结果 RT-PCR及Westen blot结果提示,FoxM1在HT-29细胞中高表达,而在HCT-116细胞中呈低表达。应用PEX-2-FoxM1上调HCT-116细胞中FoxM1表达后,细胞的划痕愈合能力HCT-116实验组[(70.92±1.48)%]较HCT-116实验对照组[(18.43±3.01)%]及HCT-116空白对照组[(16.66±2.63)%]显著增强(P<0.05)。HCT-116实验组Transwell小室穿膜细胞数[(186.0±6.8)个]较HCT-116实验对照组[(42.0±2.0)个]及HCT-116空白对照组[(37.0±2.2)个]均增加(P<0.05)。应用pGPH-sh FoxM1下调HT-29细胞中FoxM1表达后,HT-29实验组细胞的划痕愈合能力[(10.37±3.86)%]较HT-29实验对照组[(39.79±2.17)%]及HT-29空白对照组[(67.36±2.61)%]显著下降(P<0.05)。HT-29实验组Transwell小室穿膜细胞数[(53.0±1.8)个]较较HT-29实验对照组[(95.0±2.2)个]及HT-29空白对照组[(118.0±4.0)个]均减少(P<0.05)。结论FoxM1表达与结肠癌的侵袭、转移等关系密切;FoxM1的siRNA干扰可有效抑制结肠癌细胞的增殖、迁移和侵袭,人为调高结肠癌细胞株中FoxM1表达则促进细胞的增殖、迁移和侵袭。提示FoxM1可能成为抑制结肠癌细胞增殖和转移新的分子靶点。
The purpose of colon cancer invasion and metastasis is the main cause of death in patients with colon cancer. This article intends to explore the impact of Forkhead box M1 (FoxM1) on the malignant phenotype of human colon cancer cells. Methods High-expression cell line HT-29 and low-expression cell line HCT-116 were screened by real time-PCR (RT-PCR) and Western blotting. RNA interference was used to down-regulate the expression of FoxM1 in HT-29 cells While the expression of FoxM1 in HCT-116 was induced by overexpression plasmid. The two cell lines were divided into three groups according to non-transfected, empty vector and transfection interference or overexpression of FoxM1 plasmid: blank Control group, experimental control group and experimental group. Scratch assay and Transwell chamber assay were used to detect the cell proliferation, migration and invasion ability of the above transfected cells. Results RT-PCR and Westen blot suggested that FoxM1 was highly expressed in HT-29 cells and low in HCT-116 cells. HCT-116 experimental group [(70.92 ± 1.48)%] compared with HCT-116 experimental control group [(18.43 ± 3.01)%] And HCT-116 control group [(16.66 ± 2.63)%] (P <0.05). Compared with HCT-116 control group [(42.0 ± 2.0)] and HCT-116 control group [(37.0 ± 2.2)], the number of transwell cells in HCT-116 group [(186.0 ± 6.8) Increase (P <0.05). Compared with HT-29 experimental control group [(39.79 ± 2.17)%] and HT-29 experimental group (SciV), scratch healing ability of HT-29 experimental group was significantly decreased by pGPH-sh FoxM1 down-regulation of FoxM1 expression in HT- HT-29 blank control group [(67.36 ± 2.61)%] significantly decreased (P <0.05). Compared with HT-29 experimental group [(95.0 ± 2.2)] and HT-29 blank control group [(118.0 ± 4.0)], Decreased (P <0.05). Conclusion The FoxM1 expression is closely related to the invasion and metastasis of colon cancer. FoxM1 siRNA can effectively inhibit the proliferation, migration and invasion of colon cancer cells. Human FoxM1 expression in colon cancer cell lines can promote the proliferation, migration and invasion of human colon cancer cells . Tip FoxM1 may be a new molecular target of inhibiting the proliferation and metastasis of colon cancer cells.