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±展开更多
目的:制备抗干扰素诱导的跨膜蛋白-1(interferon-induced transmembrane protein 1,IFITM1)的单克隆抗体,为检测IFITM1及进一步研究其在结肠肿瘤发生过程中的作用提供实验基础。方法:以结肠癌患者的癌组织为材料,提取总RNA,以RT-PCR扩...目的:制备抗干扰素诱导的跨膜蛋白-1(interferon-induced transmembrane protein 1,IFITM1)的单克隆抗体,为检测IFITM1及进一步研究其在结肠肿瘤发生过程中的作用提供实验基础。方法:以结肠癌患者的癌组织为材料,提取总RNA,以RT-PCR扩增得到IFITM1 cDNA序列,经EcoRⅠ和HindⅢ双酶切后,克隆入pGEX-4T-3进行原核表达并纯化得IFITM1-GST;以该融合蛋白免疫BALB/c小鼠,淋巴细胞杂交瘤法制备单克隆抗体;采用ELISA、Western-blot及免疫组织化学法以制备的抗体检测结肠癌患者结肠癌组织中的IFITM1。结果:成功构建了IFITM1原核表达载体,获得了IFITM1-GST重组蛋白;制备得到了1株抗IFITM1单克隆抗体,腹水ELISA效价为1:30000,抗体亚类为IgG1,可用于ELISA、Western-blot及免疫组织化学法检测结肠癌患者结肠癌组织中的IFITM1。结论:获得了1株可用于ELISA、Western-blot及免疫组织化学法的抗IFITM1单克隆抗体2F-1,为进一步研究IFITM1在结肠肿瘤发生过程中的作用提供了实验基础。展开更多
目的:探讨干扰素介导的跨膜蛋白1(Interferon-induced transmembrane protein 1,IFITM1)基因在卵巢上皮性癌中表达的相关性及其意义。方法:应用Western blotting检测正常卵巢、卵巢良性肿瘤、卵巢交界性肿瘤和卵巢上皮性癌组织中IFITM1...目的:探讨干扰素介导的跨膜蛋白1(Interferon-induced transmembrane protein 1,IFITM1)基因在卵巢上皮性癌中表达的相关性及其意义。方法:应用Western blotting检测正常卵巢、卵巢良性肿瘤、卵巢交界性肿瘤和卵巢上皮性癌组织中IFITM1蛋白表达。免疫组织化学检测12例正常卵巢、21例卵巢良性肿瘤、18例卵巢交界性肿瘤和85例卵巢上皮性癌组织中IFITM1的蛋白表达,同时分析IFITM1表达状况与临床病理因素之间的相关性。结果:Western blotting显示卵巢上皮性癌和卵巢交界性肿瘤中IFITM1表达水平明显高于正常卵巢组织和卵巢良性肿瘤。免疫组化显示在正常卵巢组织中IFITM1阳性表达率为41.7%(5/12),在卵巢良性肿瘤组织中71.4%(15/21),在卵巢交界性肿瘤组织中为72.2%(13/18),在卵巢上皮性癌中为77.6%(66/85),IFITM1蛋白表达强度在正常卵巢、良性卵巢肿瘤、交界性卵巢肿瘤、上皮性卵巢癌间的比较有统计学意义(P<0.05)。IFITM1蛋白表达与病理类型、肿瘤分化程度、肿瘤FIGO分期有关(P<0.05),与淋巴结转移、腹水无明显相关性。化疗敏感组和耐药组的IFITM1表达强度间差异有统计学意义(P<0.05)。结论:IFITM1在正常卵巢、卵巢良性肿瘤、卵巢交界性肿瘤和卵巢上皮性癌组织中的表达依次升高,并与卵巢癌以铂类为基础的化疗耐药性产生有相关性,为进一步研究IFITM1在卵巢癌诊治及化疗中的应用前景提供依据。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±
文摘目的:制备抗干扰素诱导的跨膜蛋白-1(interferon-induced transmembrane protein 1,IFITM1)的单克隆抗体,为检测IFITM1及进一步研究其在结肠肿瘤发生过程中的作用提供实验基础。方法:以结肠癌患者的癌组织为材料,提取总RNA,以RT-PCR扩增得到IFITM1 cDNA序列,经EcoRⅠ和HindⅢ双酶切后,克隆入pGEX-4T-3进行原核表达并纯化得IFITM1-GST;以该融合蛋白免疫BALB/c小鼠,淋巴细胞杂交瘤法制备单克隆抗体;采用ELISA、Western-blot及免疫组织化学法以制备的抗体检测结肠癌患者结肠癌组织中的IFITM1。结果:成功构建了IFITM1原核表达载体,获得了IFITM1-GST重组蛋白;制备得到了1株抗IFITM1单克隆抗体,腹水ELISA效价为1:30000,抗体亚类为IgG1,可用于ELISA、Western-blot及免疫组织化学法检测结肠癌患者结肠癌组织中的IFITM1。结论:获得了1株可用于ELISA、Western-blot及免疫组织化学法的抗IFITM1单克隆抗体2F-1,为进一步研究IFITM1在结肠肿瘤发生过程中的作用提供了实验基础。
