Background Interferon-induced transmembrane protein 1 （IFITM1） has been identified as a molecular marker of the colorectal tumors; however its influences on the biological behaviors of the colorectal cancer cells ar...Background Interferon-induced transmembrane protein 1 （IFITM1） has been identified as a molecular marker of the colorectal tumors; however its influences on the biological behaviors of the colorectal cancer cells are currently unknown.We aimed to study the influences of IFITM1 on the proliferation,invasion,and metastasis of the colorectal cancer SW480 cell lines.Methods We constructed IFITM1/pEGFP-C3 recombinant plasmids and transfected them into the colorectal cancer SW480 cell lines.IFITM1/pEGFP-C3 recombinant plasmids were identified by means of immunofluorescence,laser confocal scanning microscopy,and reverse transcription polymerase chain reaction.IFITM1/SW480 cells with stable over-expression of IFITM1 were confirmed by G418 screening.The influences of IFITM1 on the proliferation of the SW480 cell lines were investigated by MTT assay and tumor transplantation experiments in nude mice.Cell invasion experiments were performed to determine the invasion capacity of the IFITM1/SW480 cells.Matrix metalloproteinase 2 （MMP-2） and MMP-9 activities were detected by the gelatin zymographic analysis,and MMP-9 expression by the Western blotting analysis.Results IFITM1/pEGFP-C3 recombinant plasmids were successfully constructed in this study,and the IFITM1/SW480 cells with stable IFITM1 gene over-expression were confirmed by G418 screening.MTT results showed that the proliferation of the IFITM1/SW480 cells was significantly enhanced （P 〈0.01）.Tumors were harvested from four weeks old mice.Tumor volumes were （1347.00±60.94） mm3,（1032.40±111.38） mm3 and （1018.78±28.83） mm3; and tumor weights were （1522.34±62.76） mg,（1137.78±97.22） mg and （1155.76±133.31） mg for mice inoculated with the IFITM1/SW480 cells,pEGFP-C3/SW480 cells and SW480 cells,respectively.Tumor volumes and weights from mice inoculated with the IFITM1/SW480 cells were significantly increased （P 〈0.01）.In addition,the numbers of the SW480 cells and IFITM1/SW480 cells that migrated through Matrigel were 448.64±38.09 and 540.45±44.61,respectively; so the invasive ability of the SW480 cells transfected with IFITM1 gene was significantly greater than that of the SW480 cells （P 〈0.01）.Gelatin zymographic analysis showed that MMP-9 and MMP-2 protein activities in the IFITM1/SW480 cells were significantly enhanced,and Western blotting analysis showed that MMP-9 expression in the IFITM1/SW480 cells was also increased.Conclusion IFITMl can enhance the proliferation,invasion,and metastasis of the colorectal cancer SW480 cell lineS.展开更多
目的探讨过表达干扰素诱导跨膜蛋白3(IFITM3)对胆囊癌细胞MHCC97H增殖、迁移、侵袭的影响。方法构建IFITM3-pc DNA3.1过表达载体,并将IFITM3-pc DNA3.1及pc DNA3.1转入MHCC97H细胞中,同时设立正常对照组。采用实时荧光定量聚合酶链反应(...目的探讨过表达干扰素诱导跨膜蛋白3(IFITM3)对胆囊癌细胞MHCC97H增殖、迁移、侵袭的影响。方法构建IFITM3-pc DNA3.1过表达载体,并将IFITM3-pc DNA3.1及pc DNA3.1转入MHCC97H细胞中,同时设立正常对照组。采用实时荧光定量聚合酶链反应(PCR)法检测IFITM3、B细胞淋巴瘤/白血病-2相关X蛋白(BAX)、B细胞淋巴瘤/白血病-2(Bcl-2)表达,CCK-8法检测细胞活力,划痕实验检测细胞迁移能力,Transwell实验检测细胞侵袭能力。结果酶切结果证实IFITM3过表达载体构建成功。与正常对照组及过表达对照组比较,过表达组MHCC97H细胞IFITM3 m RNA表达量上调,光密度值(OD)升高,细胞迁移速率加快,细胞侵袭数目增加,Bcl-2 m RNA表达水平上调,BAX m RNA表达水平下调,差异均有统计学意义(P﹤0.01)。结论过表达IFITM3能显著促进MHCC97H胆囊癌细胞增殖、迁移及侵袭,与调控Bcl-2及BAX表达有关。展开更多
文摘Background Interferon-induced transmembrane protein 1 （IFITM1） has been identified as a molecular marker of the colorectal tumors; however its influences on the biological behaviors of the colorectal cancer cells are currently unknown.We aimed to study the influences of IFITM1 on the proliferation,invasion,and metastasis of the colorectal cancer SW480 cell lines.Methods We constructed IFITM1/pEGFP-C3 recombinant plasmids and transfected them into the colorectal cancer SW480 cell lines.IFITM1/pEGFP-C3 recombinant plasmids were identified by means of immunofluorescence,laser confocal scanning microscopy,and reverse transcription polymerase chain reaction.IFITM1/SW480 cells with stable over-expression of IFITM1 were confirmed by G418 screening.The influences of IFITM1 on the proliferation of the SW480 cell lines were investigated by MTT assay and tumor transplantation experiments in nude mice.Cell invasion experiments were performed to determine the invasion capacity of the IFITM1/SW480 cells.Matrix metalloproteinase 2 （MMP-2） and MMP-9 activities were detected by the gelatin zymographic analysis,and MMP-9 expression by the Western blotting analysis.Results IFITM1/pEGFP-C3 recombinant plasmids were successfully constructed in this study,and the IFITM1/SW480 cells with stable IFITM1 gene over-expression were confirmed by G418 screening.MTT results showed that the proliferation of the IFITM1/SW480 cells was significantly enhanced （P 〈0.01）.Tumors were harvested from four weeks old mice.Tumor volumes were （1347.00±60.94） mm3,（1032.40±111.38） mm3 and （1018.78±28.83） mm3; and tumor weights were （1522.34±62.76） mg,（1137.78±97.22） mg and （1155.76±133.31） mg for mice inoculated with the IFITM1/SW480 cells,pEGFP-C3/SW480 cells and SW480 cells,respectively.Tumor volumes and weights from mice inoculated with the IFITM1/SW480 cells were significantly increased （P 〈0.01）.In addition,the numbers of the SW480 cells and IFITM1/SW480 cells that migrated through Matrigel were 448.64±38.09 and 540.45±44.61,respectively; so the invasive ability of the SW480 cells transfected with IFITM1 gene was significantly greater than that of the SW480 cells （P 〈0.01）.Gelatin zymographic analysis showed that MMP-9 and MMP-2 protein activities in the IFITM1/SW480 cells were significantly enhanced,and Western blotting analysis showed that MMP-9 expression in the IFITM1/SW480 cells was also increased.Conclusion IFITMl can enhance the proliferation,invasion,and metastasis of the colorectal cancer SW480 cell lineS.
文摘目的探讨过表达干扰素诱导跨膜蛋白3(IFITM3)对胆囊癌细胞MHCC97H增殖、迁移、侵袭的影响。方法构建IFITM3-pc DNA3.1过表达载体,并将IFITM3-pc DNA3.1及pc DNA3.1转入MHCC97H细胞中,同时设立正常对照组。采用实时荧光定量聚合酶链反应(PCR)法检测IFITM3、B细胞淋巴瘤/白血病-2相关X蛋白(BAX)、B细胞淋巴瘤/白血病-2(Bcl-2)表达,CCK-8法检测细胞活力,划痕实验检测细胞迁移能力,Transwell实验检测细胞侵袭能力。结果酶切结果证实IFITM3过表达载体构建成功。与正常对照组及过表达对照组比较,过表达组MHCC97H细胞IFITM3 m RNA表达量上调,光密度值(OD)升高,细胞迁移速率加快,细胞侵袭数目增加,Bcl-2 m RNA表达水平上调,BAX m RNA表达水平下调,差异均有统计学意义(P﹤0.01)。结论过表达IFITM3能显著促进MHCC97H胆囊癌细胞增殖、迁移及侵袭,与调控Bcl-2及BAX表达有关。
